The membrane-associated c-type cytochromes(c-Cyts) have been well known as the key enzymes mediating extracellular electron transfer to terminal electron acceptors, resulting in biogeochemical elemental transformation...The membrane-associated c-type cytochromes(c-Cyts) have been well known as the key enzymes mediating extracellular electron transfer to terminal electron acceptors, resulting in biogeochemical elemental transformation, contaminant degradation, and nutrient cycling. Although c-Cyts-mediated metal reduction or oxidation have been mainly investigated with the purified proteins of metal reducing/oxidizing bacteria, the in vivo behavior of c-Cyts is still unclear, given the difficulty in measuring the proteins of intact cells. Fortunately, the in situ spectroscopy would be ideal for measuring the reaction kinetics of c-Cyts in intact cells under noninvasive physiological conditions. It can also help the establishment of kinetic/thermodynamic models of extracellular electron transfer processes, which are essential to understand the electron transfer mechanisms at the molecular scale. This review briefly summarizes the current advances in spectral methods for examining the c-Cyts in intact cells of dissimilatory metal reducing bacteria and Fe(Ⅱ)-oxidizing bacteria.展开更多
Cytochrome c(cyt c)is released from mitochondria into the cytosol upon apoptotic stimulation,ultimately triggering programmed cell death.Recent studies have revealed that transfer RNA(tRNA)interacts with cyt c,impedin...Cytochrome c(cyt c)is released from mitochondria into the cytosol upon apoptotic stimulation,ultimately triggering programmed cell death.Recent studies have revealed that transfer RNA(tRNA)interacts with cyt c,impeding the formation of the apoptosome complex and thereby suppressing apoptosis.To elucidate the molecular mechanism underlying the interaction between cyt c and tRNA,nuclear magnetic resonance(NMR)-based chemical shift perturbation and intensity analysis were employed to characterize the binding interface between cyt c and tRNAphe.The findings demonstrate that cyt c primarily engages with tRNAphe through its 70–85Ω-loop and N-terminalα-helix.This interaction sterically hinders the accessibility of small molecules,such as H_(2)O_(2),to the hydrophobic pocket of cyt c,consequently attenuating its peroxidase activity.Furthermore,oxidative modification of cyt c,particularly the carbonylation of positively charged lysine residues,weakens this interaction.展开更多
Regulation of apoptosis represents a key parameter in all living organisms.In this paper,an input-induced logic-gated modular nanocalculator is designed to regulate cancer cell apoptosis by programmatically combining ...Regulation of apoptosis represents a key parameter in all living organisms.In this paper,an input-induced logic-gated modular nanocalculator is designed to regulate cancer cell apoptosis by programmatically combining and connecting logic gate modules with different functions.Via rational design of the various logic gate modules of the nanocalculator,different apoptosis related operations including cancer cell targeting,apoptosis induction,and apoptosis monitoring could be performed.Importantly,each of these logic gate modules could independently perform apoptosis related YES logic operations when ran separately.After combining each YES logic gate module into a logic circuit and connecting it to the GO scaffold to construct a logic-gated nanocalculator,the input-induced logic-gated modular nanocalculator could selectively enter cancer cells and control the drug release to logically apoptosis(output),by performing AND logic gate operations when inputs(nucleolin and H^(+)) were included at the same time.Moreover,evidence suggests that these efficient logical calculations proceed in cancer cell apoptosis regulation without the general limiations of lithography in nanotechnology.As such,this work provides a new vision for the construction of a logic-gated modular nanocalculator with logical calculation proficiency potentially useful in cancer therapy and the regulation of life.展开更多
Many labdane-related diterpenoids(LRDs) exhibit high values in drug development.Their diversity in structure and bioactivity,to a large extent,arise from oxidative modifications which are mainly catalyzed by cytochrom...Many labdane-related diterpenoids(LRDs) exhibit high values in drug development.Their diversity in structure and bioactivity,to a large extent,arise from oxidative modifications which are mainly catalyzed by cytochrome P450s(CYPs).The medicinal plant Euphorbia fischeriana Steud.is rich in LRDs with distinct scaffolds.Herein,we characterized three cytochrome P450s involved in LRD biosynthesis from this plant.Notably,CYP71D450 and CYP701A148 are two substrate-promiscuity CYPs.The former is the first example of CYPs which can oxidize C-3 of ent-atisane skeleton and ent-isopimara-7(8),15-diene,and the latter is the first example of CYPs which can oxidize C-19 of ent-abietane and ent-pimarane skeletons.This study expands the toolkit for bioproduction of diverse LRDs.展开更多
Background:A significant proportion of patients still cannot benefit from existing targeted therapies and immunotherapies,making the search for new treatment strategies extremely urgent.In this study,we combined integ...Background:A significant proportion of patients still cannot benefit from existing targeted therapies and immunotherapies,making the search for new treatment strategies extremely urgent.In this study,we combined integrate public data analysis with experimental validation to identify novel prognostic biomarkers and therapeutic targets for lung adenocarcinoma(LUAD).Methods:We analyzed RNA and protein databases to assess the expression levels of cytochrome C oxidase 5B(COX5B)in LUAD.Several computational algorithms were employed to investigate the relationship between COX5B and immune infiltration in LUAD.To further elucidate the role of COX5B in LUAD,we utilized multiple experimental approaches,including quantitative reverse transcription PCR assays,western blot,immunohistochemistry,electron microscopy,flow cytometry,and EdU proliferation assays.Results:We revealed that COX5B was significantly elevated in LUAD and positively correlated with poor prognosis of LUAD patients.Analysis of co-expression network indicated that COX5B may take part in the intracellular adenosine triphosphate(ATP)synthesis through the oxidative phosphorylation pathway.There was a negative correlation between COX5B expression and immune infiltration in LUAD.Furthermore,we validated that COX5B levels were significantly elevated in both LUAD tissues and cell lines.Specifically,immunohistochemistry(IHC)assays revealed a 2.32-fold increase of COX5B in tumor tissues compared to that in adjacent normal tissues(p=0.0044).Additionally,COX5B knockdown disrupted the redox homeostasis,ultimately suppressed the proliferation of LUAD cells.Subsequent investigations demonstrated that berberine effectively targeted COX5B,diminishing its protein expression and consequently inhibiting cell proliferation and tumor growth in LUAD.Conclusions:This study established that upregulated COX5B was positive associated with poor patient prognosis in LUAD,elucidating the mechanisms by which berberine targets COX5B to inhibit tumor growth,thereby providing a novel therapeutic target and strategy for the clinical management of LUAD.展开更多
The Saccostrea mordax Gould,1850 is a typical intertidal species,whose genetic differentiation is influenced by various factors,including geological and climatic changes.To explore the genetic structure and historical...The Saccostrea mordax Gould,1850 is a typical intertidal species,whose genetic differentiation is influenced by various factors,including geological and climatic changes.To explore the genetic structure and historical population characteristics of Saccostrea mordax,we sequenced the mitochondrial cytochrome c oxidase subunit I(COI)gene from 58 specimens sampled from four locations in the western Pacific.Additionally,103 individuals from the Persian Gulf and western Pacific(from databases)were included for phylogenetic analysis.The Bayesian Inference tree showed that all specimens were divided into two clades,i.e.,the Persian Gulf population and the western Pacific population.Spatial molecular variance analysis indicated significant genetic differentiation between the two populations,and isolation by distance analysis revealed a positive correlation between genetic differentiation and geographic distance.Neutrality tests and Bayesian Skyline Plot suggested that both populations underwent expansions during the late Pleistocene.This study revealed the population history of Saccostrea mordax and described a new lineage,Saccostrea mordax lineage D,providing a foundation for understanding oyster biodiversity formation and genetic resource conservation.展开更多
D-a-Tocopheryl polyethylene glycol 1000 succinate(TPGS,also known as vitamin E-TPGS)is a biodegradable amphiphilic polymer prepared by esterification of vitamin E with polyethylene glycol(PEG)1000.It is approved by th...D-a-Tocopheryl polyethylene glycol 1000 succinate(TPGS,also known as vitamin E-TPGS)is a biodegradable amphiphilic polymer prepared by esterification of vitamin E with polyethylene glycol(PEG)1000.It is approved by the US Food and Drug Administration(FDA)and has found wide application in nanocarrier drug delivery systems(NDDS).Fully characterizing the in vivo fate and pharmacokinetic behavior of TPGS is important to promote the further development of TPGS-based NDDS.However,to date,a bioassay for the simultaneous quantitation of TPGS and its metabolite,PEG1000,has not been reported.In the present study,we developed such an innovative bioassay and used it to investigate the pharmacokinetics,tissue distribution and excretion of TPGS and PEG1000 in rat after oral and intravenous dosing.In addition,we evaluated the interaction of TPGS with cytochromes P450(CYP450s)in human liver microsomes.The results show that TPGS is poorly absorbed after oral administration with very low bioavailability and that,after intravenous administration,TPGS and PEG1000 are mainly distributed to the spleen,liver,lung and kidney before both being slowly eliminated in urine and feces as PEG1000.In vitro studies show the inhibition of human CYP450 enzymes by TPGS is limited to a weak inhibition of CYP3A4.Overall,our results provide a clear picture of the in vivo fate of TPGS which will be useful in evaluating the safety of TPGS-based NDDS in clinical use and in promoting their further development.展开更多
Direct interspecies electron transfer(DIET)may be most important in methanogenic environments,but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron ac...Direct interspecies electron transfer(DIET)may be most important in methanogenic environments,but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron acceptor.To better understand DIET with methanogens,the transcriptome of Geobacter metallireducens during DIET‐based growth with G.sulfurreducens reducing fumarate was compared with G.metallireducens grown in coculture with diverse Methanosarcina.The transcriptome of G.metallireducens cocultured with G.sulfurreducens was significantly different from those with Methanosarcina.Furthermore,the transcriptome of G.metallireducens grown with Methanosarcina barkeri,which lacks outer‐surface c‐type cytochromes,differed from those of G.metallireducens cocultured with M.acetivorans or M.subterranea,which have an outer‐surface c‐type cytochrome that serves as an electrical connect for DIET.Differences in G.metallireducens expression patterns for genes involved in extracellular electron transfer were particularly notable.Cocultures with c‐type cytochrome deletion mutant strains,ΔGmet_0930,ΔGmet_0557 andΔGmet_2896,never became established with G.sulfurreducens but adapted to grow with all three Methanosarcina.Two porin–cytochrome complexes,PccF and PccG,were important for DIET;however,PccG was more important for growth with Methanosarcina.Unlike cocultures with G.sulfurreducens and M.acetivorans,electrically conductive pili were not needed for growth with M.barkeri.Shewanella oneidensis,another electroactive microbe with abundant outer‐surface c‐type cytochromes,did not grow via DIET.The results demonstrate that the presence of outer‐surface c‐type cytochromes does not necessarily confer the capacity for DIET and emphasize the impact of the electron‐accepting partner on the physiology of the electron‐donating DIET partner.展开更多
P450 enzymes-catalyzed aromatic hydroxylation plays an important role in detoxification,biosynthesis,and potential carcinogenic effect of aromatic compounds.Though it has been explored for decades,the actual process o...P450 enzymes-catalyzed aromatic hydroxylation plays an important role in detoxification,biosynthesis,and potential carcinogenic effect of aromatic compounds.Though it has been explored for decades,the actual process of aromatic hydroxylation and mechanism of regioselectivity catalyzed by cytochrome P450 monooxygenases remained ambiguous.Here,we have resolved these issues.With a stable chiral organofluorine probe,and especially with X-ray data of two isolated arene oxides derivatives,we demonstrate that an arene oxide pathway is definitely involved in P450-catalyzed aromatic hydroxylation.By the capture,isolation,identification and reactivity exploration of the arene 1,2-oxide and arene 2,3-oxide intermediates,together with advanced QM calculations,the mechanism of how two intermediates go to the same product has been elucidated.In addition to the model substrate,we also confirmed that an arene oxide intermediate is involved in the P450-catalyzed hydroxylation pathway of a natural product derivative methyl cinnamate,which indicates that this intermediate appears to be universal in P450-catalyzed aromatic hydroxylation.Our work not only provides the most direct evidence for the arene oxide pathway and new insights into the regioselectivity involved in P450-catalyzed aromatic hydroxylation,but also supplies a new synthetic approach to achieve the dearomatization of aromatic compounds.展开更多
Rice sheath blight(RSB)is a major destructive disease impeding rice production.Identifying key germplasm resources with increased resistance remains a challenge.However,the mechanisms underlying disease resistance are...Rice sheath blight(RSB)is a major destructive disease impeding rice production.Identifying key germplasm resources with increased resistance remains a challenge.However,the mechanisms underlying disease resistance are not yet fully understood.Cytochrome P450 monooxygenases(CYP450s)serve biosynthesis and metabolic detoxification functions in plants,but there is limited information about their role in the response induced by RSB.This study demonstrated that CYT02 belongs to the CYP73A100 subfamily and is a typical member of the CYP450s.Overexpression(OE)in rice of the cytochrome P450 monooxygenase cyt02 conferred increased resistance to RSB and increased vegetative tillering.Cyt02 may increase RSB resistance by regulating plant hormone synthesis,regulate reactive oxygen species(ROS)by coordinating the activity of antioxidant enzymes,and initiate phytoalexin synthesis in response to fungal infection.These research findings have laid a foundation for a deeper understanding of the function of cyt02 and offered a potential target gene for breeding rice varieties resistant to sheath blight.展开更多
Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration ...Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Iα-loop-helix Iβmotif which corresponds to helix I of other P 450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,theγ-domain.The structure reveals a homodimer in which theγ-domain of one molecule interacts with theβ-domain of another.The plane of heme group makes an angle of approximately 40°with the helix Iα-loop-helix Iβmotif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recognition.Phe-301,Ser-303 and Gly-305 from the helix Iα-loop-helix Iβmotif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.展开更多
A class of persistent organic pollutants,polychlorinated biphenyls(PCBs),are ubiquitous in the environment and human tissues which are continuously and long-term threatened.We aim to investigate the toxic effects and ...A class of persistent organic pollutants,polychlorinated biphenyls(PCBs),are ubiquitous in the environment and human tissues which are continuously and long-term threatened.We aim to investigate the toxic effects and the underlying mechanisms of PCB153 using Caenorhabditis elegans as a model organism.Our findings demonstrated that exposure to an optimized concentration of 2μmol/L PCB153 had adverse effects on C.elegans,led to reduction of lifespan,body length,frequency of body bending,and head wiggling.Additionally,this exposure led to an increase in the accumulation of reactive oxygen species,superoxide dismutase,lipofuscin and fat content within the organism.Furthermore,gene set enrichment analysis and whole transcriptome sequencing revealed,a significant up-regulation of Cytochrome P450(CYP)family genes,which are crucial for the xenobiotics metabolism in C.elegans.Knocking down specific genes within the CYPs family,including cyp-35C1,cyp-35A5,cyp-35A3,cyp-34A1,and cyp-34A10,via RNA interference were able to reverse the shortening of lifespan and fat accumulation induced by PCB153 exposure.Moreover,a comprehensive a competing endogenous RNA network was constructed by integrating qPCR-validated differentially expressed m RNA along with lncRNA,circRNA,and miRNA identified by transcriptomic sequencing.This study emphasizes that PCB153 exposure leads to the physiological impairments,particularly fat accumulation and a shorter lifespan,through the modulation of CYP450 family genes.These findings contribute to our understanding of environmental pollutants and their impact on biological systems and provide valuable information on the toxic effects and potential risks associated with PCB153 exposure.展开更多
A comprehensive taxonomic reassessment of the Hebius optatus species complex,integrating molecular data and morphological data,has led to the recognition and description of a previously overlooked species from norther...A comprehensive taxonomic reassessment of the Hebius optatus species complex,integrating molecular data and morphological data,has led to the recognition and description of a previously overlooked species from northern Vietnam and the Guangxi Zhuang Autonomous Region,China,herein described as Hebius vogeli sp.nov.Phylogenetic analyses reveal that Hebius vogeli sp.nov.is closely related to H.sauteri and H.optatus sensu stricto.Uncorrected p-distances based on cytochrome b sequences indicate a divergence of 10.74%-11.73%between Hebius vogeli sp.nov.and H.optatus.The new species differs from H.optatus by a combination of characters,including a higher number of subcaudal scales,a greater total number of ventral and subcaudal scales,and distinct ventral coloration.Hebius vogeli sp.nov.appears to be geographically restricted to northern Vietnam and the adjacent region of Guangxi Zhuang Autonomous Region,China,making its distribution allopatric from that of H.optatus.The Pearl River is hypothesized to serve as a biogeographic barrier separating the two taxa.Accordingly,we recommend the exclusion of H.optatus from the herpetofauna of Vietnam.Based on IUCN Red List criteria,we propose that Hebius vogeli sp.nov.be assessed as Least Concern(LC).Our results emphasize both the underestimated diversity within Hebius and the need for continued integrative taxonomic efforts,particularly in the Vietnam-China borderlands,where cryptic diversity remains largely unexplored.展开更多
BACKGROUND Diabetic nephropathy(DN)is a major complication of diabetes,marked by progressive renal damage and an inflammatory response.Although research has investigated the pathological mechanisms underlying DN,effec...BACKGROUND Diabetic nephropathy(DN)is a major complication of diabetes,marked by progressive renal damage and an inflammatory response.Although research has investigated the pathological mechanisms underlying DN,effective treatment options remain limited.AIM To evaluate the therapeutic impact of Shenzhuo formulation(SZF)on a DN mouse model and to examine its potential molecular mechanisms using transcriptomic and metabolomic approaches.METHODS We established a DN mouse model through a high-fat diet combined with streptozotocin(STZ)injection,followed by SZF treatment.We analyzed SZF’s effects on gene expression and metabolite profiles in renal tissues of DN mice using transcriptomics and metabolomics techniques.Additionally,based on transcriptomic and non-targeted metabolomic findings,we further assessed SZF’s influence on the expression of factors related to the cytochrome P450(CYP450)-mediated arachidonic acid(AA)metabolism pathway,as well as its effects on inflammation and oxidative stress.RESULTS SZF intervention significantly decreased hyperglycemia and mitigated renal function impairment in DN mice.Pathological analysis revealed that SZF treatment improved renal tissue damage,reduced fibrosis,and diminished glycogen deposition.Transcriptomic analysis indicated that SZF influenced mRNA expression of CYP450-related genes,including Cyp2j13,Cyp2b9,Pla2 g2e/Cyp4a12a,Cyp4a32,Cyp2e1,and Cyp4a14.Non-targeted metabolomic results demonstrated that SZF altered the levels of metabolites associated with the AA metabolic pathway,including 5,6-EET,14,15-EET,phosphatidylcholine,and 20-HETE.Further experiments showed that SZF upregulated the expression of CYP4A and CYP2E proteins in renal tissue,as well as CYP2J and CYP2B proteins.Additionally,SZF significantly reduced the expression of inflammatory factors in renal tissue,enhanced antioxidant enzyme activity,and alleviated oxidative stress.CONCLUSION SZF exerts anti-inflammatory and antioxidant effects by regulating CYP450-mediated AA metabolism,leading to improved renal function and improved pathological state in DN mice.展开更多
The cytochrome P4503A(CYP3A)gene family’s role in early progression of gastric cancer was comprehensively investigated.Its potential as a therapeutic target was evaluated.Upon literature review,aberrant expression of...The cytochrome P4503A(CYP3A)gene family’s role in early progression of gastric cancer was comprehensively investigated.Its potential as a therapeutic target was evaluated.Upon literature review,aberrant expression of the CYP3A gene family has a strong correlation with gastric cancer onset,although the precise underlying mechanisms remain unclear.To assess its potential as a biomarker for early diagnosis and a therapeutic target,we have provided a comprehensive review of the regulatory mechanisms governing CYP3A gene family expression in gastric cancer,as well as its relation with early tumor progression and the tumor microenvironment.The CYP3A gene family is crucial in the proliferation,migration,and invasion of gastric cancer cells and promotes cancer progression by modulating inflammatory responses and oxidative stress within the tumor microenvironment.Furthermore,genetic polymorphisms in CYP3A enzymes highlight its potential value in personalized medicine.Based on these findings,this paper explores the feasibility of developing inhibitors and activators targeting CYP3A enzymes and discusses potential applications in gene therapy.This research provides crucial theoretical support for the CYP3A gene family as an early diagnostic marker and therapeutic target for gastric cancer.In the future,multi-omics studies and large-scale clinical trials will be essential to advance clinical translation of these findings.展开更多
Heme peroxygenases exhibit remarkable catalytic versatility in facilitating a wide array of oxidative reactions under mild conditions,eliminating the need for coenzymes and intricate electron transport systems.This un...Heme peroxygenases exhibit remarkable catalytic versatility in facilitating a wide array of oxidative reactions under mild conditions,eliminating the need for coenzymes and intricate electron transport systems.This unique character underscores their essentiality and potential as promising tools in synthetic biology.Recent advancements in enzyme engineering have significantly enhanced the catalytic performance of both natural and artificial peroxygenases.Extensive engineering efforts have been directed towards unspecific peroxygenases and fatty acid peroxygenases,aiming to expand their substrate specificities,and enhance reaction selectivities,as well as increase enzyme stability.Furthermore,innovative strategies such as dual-functional small molecule-assisted systems and H_(2)O_(2) tunnel engineering have been harnessed to transform P450 monooxygenases into highly efficient peroxygenases,capable of catalyzing reactions with a variety of unnatural substrates.This review consolidates the latest progress in the engineered and artificial heme peroxygenases,emphasizing their catalytic performances as potent biocatalysts for sustainable organic synthesis.展开更多
The tobacco whitefly,Bemisia tabaci,is a notorious pest affecting various crops globally,and it exhibits high levels of resistance to various insecticides.Afidopyropen is a recently commercialized pyropene insecticide...The tobacco whitefly,Bemisia tabaci,is a notorious pest affecting various crops globally,and it exhibits high levels of resistance to various insecticides.Afidopyropen is a recently commercialized pyropene insecticide for B.tabaci control with high selectivity and a novel mode of action.We previously identified a high level of afidopyropen resistance in a field-collected population after selection in the lab,and named it the HD-Afi strain.In the present study,minimal cross-resistance in the HD-Afi strain was found between afidopyropen and other common chemical agents.However,the P450 enzyme activity in HD-Afi was 2.18 times the level in susceptible strain HD-S.Expression analysis revealed that two of 12 candidate P450 genes,namely CYP6DW3 and CYP4C64,were significantly up-regulated in HD-Afi.Silencing CYP6DW3 and CYP4C64 by RNA interference(RNAi)substantially increased the susceptibility of whitefly adults,confirming their involvement in afidopyropen resistance.Homology modeling and molecular docking analyses demonstrated stable binding of afidopyropen to CYP6DW3 and CYP4C64,with binding free energies of–6.87 and–6.11 kcal mol^(-1),respectively.The findings of this study suggest that the induction of CYP6DW3 and CYP4C64 facilitates afidopyropen detoxification,contributing to the development of resistance in B.tabaci.展开更多
Background:The thermogenic characteristics of animals are closely related to species distribution,and basal metabolic rate(BMR)and thermal neutral zone(TNZ)are important components of the thermogenic process in animal...Background:The thermogenic characteristics of animals are closely related to species distribution,and basal metabolic rate(BMR)and thermal neutral zone(TNZ)are important components of the thermogenic process in animals.Furthermore,the cytochrome c oxidase 1(COX1)gene has become a subject of particular interest due to its high degree of sequence conservation,stable evolutionary rate,and rare insertions/deletions.Method:The present study selected 29 species of Rodentia and 20 species of Chiroptera.The present study employed the statistical software SPSS(27.0 Chinese version)and Origin(2024 Chinese version)software to conduct correlation analyses on a variety of biological and ecological variables.These variables included body weight,BMR,TNZ,upper thermal neutral zone,lower thermal neutral zone(LTNZ),litter size,and dietary patterns of species from different latitudes.Furthermore,we conducted a series of phylogenetic tree analyses on COX1 protein.Results:As the geographic location of a species increases in latitude,there is an observed upward trend in both body mass and BMR of rodent species.Rodentia and Chiroptera species have been observed to exhibit a decrease in LTNZ and an expansion of TNZ.With regard to dietary habits,Rodentia species are predominantly phytophagous or omnivorous.Omnivorous species exhibit a marked tendency to produce larger litters in comparison to their herbivorous counterparts.Chiroptera species exhibit a diverse dietary range,including phytophagous,carnivorous,and omnivorous species.However,no correlation was observed between dietary differences and litter size.Phylogenetic analysis of the COX1 protein subsequently demonstrated that these two species groups share a monophyletic origin.Conclusion:The present study suggests that the selected Rodentia and Chiroptera species adapt to high-latitude environments by lowering the LTNZ and widening the TNZ.Furthermore,an upward trend has been observed in the body mass and BMR of high-latitude Rodentia.Phylogenetic analysis of the COX1 protein across various taxonomic groups substantiates the efficacy of this gene for species identification.Integrating physiological phenotypes with COX1 protein molecular evidence,this study provides a reference framework for the multidimensional mechanisms of mammalian latitudinal adaptation.展开更多
The high content of cyanogenic glycosides(CG)in cassava tubers affects food safety.CG are involved in the plant growth and development and protect cassava leaves from herbivorous predators.However,the regulatory mecha...The high content of cyanogenic glycosides(CG)in cassava tubers affects food safety.CG are involved in the plant growth and development and protect cassava leaves from herbivorous predators.However,the regulatory mechanism of CG biosynthesis remains poorly understood.Here,yeast one-hybrid assays were performed using a mixed cDNA library of cassava tubers and leaves as prey and the promoter of MeCYP79D2 as bait.MeCYP79D2,a cytochrome P450 protein,is the rate-limiting enzyme for CG synthesis in cassava.From this information,a candidate regulator of MeCYP79D2 was selected and identified as transcription factor MePHD1.2.MePHD1.2,located in the nucleus and exhibiting an inhibitory transcription activity directly bound to an AT-rich motif in the promoter of MeCYP79D2.In cassava,the transcriptional activity of MeCYP79D2 was considerably enhanced in mephd1.2 mutant lines leading to increased linamarin and lotaustralin contents.Deletion of MePHD1.2 promoted the production of CGs in cassava and decreased transcription inhibition on MeCYP79D2,exposing a novel regulatory module governing biosynthesis of CGs.展开更多
Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researc...Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches.CYP80B,a family of cytochrome P450(CYP450)enzymes,plays a crucial role in BIA biosynthesis.Previously reported CYP80Bs are known to catalyze the 3′-hydroxylation of(S)-Nmethylcoclaurine,with the N-methyl group essential for catalytic activity.In this study,we successfully cloned a full-length CYP80B gene(St CYP80B)from Stephania tetrandra(S.tetrandra)and identified its function using a yeast heterologous expression system.Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that St CYP80B could catalyze Nmethylcoclaurine and coclaurine into their respective 3'-hydroxylated products.Notably,St CYP80B exhibited an expanded substrate selectivity compared to previously reported wildtype CYP80Bs,as it did not require an N-methyl group for hydroxylase activity.Furthermore,St CYP80B displayed a clear preference for the(S)-configuration.Co-expression of St CYP80B with the CYP450 reductases(CPRs,StCPR1,and StCPR2),also cloned from S.tetrandra,significantly enhanced the catalytic activity towards(S)-coclaurine.Site-directed mutagenesis of St CYP80B revealed that the residue H205 is crucial for coclaurine catalysis.Additionally,St CYP80B exhibited tissue-specific expression in plants.This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.展开更多
基金funded by the National Natural Science Foundations of China(41522105 and 41571130052)Guangdong Natural Science Funds for Distinguished Young Scholar(2014A030306041)Special Support Program(2016)
文摘The membrane-associated c-type cytochromes(c-Cyts) have been well known as the key enzymes mediating extracellular electron transfer to terminal electron acceptors, resulting in biogeochemical elemental transformation, contaminant degradation, and nutrient cycling. Although c-Cyts-mediated metal reduction or oxidation have been mainly investigated with the purified proteins of metal reducing/oxidizing bacteria, the in vivo behavior of c-Cyts is still unclear, given the difficulty in measuring the proteins of intact cells. Fortunately, the in situ spectroscopy would be ideal for measuring the reaction kinetics of c-Cyts in intact cells under noninvasive physiological conditions. It can also help the establishment of kinetic/thermodynamic models of extracellular electron transfer processes, which are essential to understand the electron transfer mechanisms at the molecular scale. This review briefly summarizes the current advances in spectral methods for examining the c-Cyts in intact cells of dissimilatory metal reducing bacteria and Fe(Ⅱ)-oxidizing bacteria.
基金financial support from National Key R&D Program of China(2018YFA0704002,2018YFE0202300,2023YFA1607500)National Natural Science Foundation of China(22174152,21991081,2204167,21505153,21675170,2147514621735007,and 22204167)+2 种基金Hubei Provincial Natural Science Foundation of China(2023AFA041)Strategic Priority Research Program of the Chinese Academy of Sciences(XDB0540300)Funding of Wuhan Special Project for Knowledge Innovation(2023020201010085).
文摘Cytochrome c(cyt c)is released from mitochondria into the cytosol upon apoptotic stimulation,ultimately triggering programmed cell death.Recent studies have revealed that transfer RNA(tRNA)interacts with cyt c,impeding the formation of the apoptosome complex and thereby suppressing apoptosis.To elucidate the molecular mechanism underlying the interaction between cyt c and tRNA,nuclear magnetic resonance(NMR)-based chemical shift perturbation and intensity analysis were employed to characterize the binding interface between cyt c and tRNAphe.The findings demonstrate that cyt c primarily engages with tRNAphe through its 70–85Ω-loop and N-terminalα-helix.This interaction sterically hinders the accessibility of small molecules,such as H_(2)O_(2),to the hydrophobic pocket of cyt c,consequently attenuating its peroxidase activity.Furthermore,oxidative modification of cyt c,particularly the carbonylation of positively charged lysine residues,weakens this interaction.
基金financially supported by the National Natural Science Foundation of China (NSFC,Nos.22134005 and 22074124)Chongqing Talents Program for Outstanding Scientists (No.cstc2021ycjh-bgzxm0178)+1 种基金Natural Science Foundation of Chongqing (No.CSTB2022NSCQ-MSX0521)the Chongqing Graduate Student Scientific Research Innovation Project (No.CYB21119)。
文摘Regulation of apoptosis represents a key parameter in all living organisms.In this paper,an input-induced logic-gated modular nanocalculator is designed to regulate cancer cell apoptosis by programmatically combining and connecting logic gate modules with different functions.Via rational design of the various logic gate modules of the nanocalculator,different apoptosis related operations including cancer cell targeting,apoptosis induction,and apoptosis monitoring could be performed.Importantly,each of these logic gate modules could independently perform apoptosis related YES logic operations when ran separately.After combining each YES logic gate module into a logic circuit and connecting it to the GO scaffold to construct a logic-gated nanocalculator,the input-induced logic-gated modular nanocalculator could selectively enter cancer cells and control the drug release to logically apoptosis(output),by performing AND logic gate operations when inputs(nucleolin and H^(+)) were included at the same time.Moreover,evidence suggests that these efficient logical calculations proceed in cancer cell apoptosis regulation without the general limiations of lithography in nanotechnology.As such,this work provides a new vision for the construction of a logic-gated modular nanocalculator with logical calculation proficiency potentially useful in cancer therapy and the regulation of life.
基金supported by grants from the National Natural Science Foundation of China [Nos.82474024,82293682 (82293680),82073953]the Nonprofit Central Research Institute Fund of Chinese Academy of Medical Sciences (No.2021-RC350-009)+1 种基金the CAMS Innovation Fund for Medical Sciences (No.2023-I2M-2-006)the Special Fund for the Construction of Qinghai Innovation Platform (No.2024-ZJ-T02)。
文摘Many labdane-related diterpenoids(LRDs) exhibit high values in drug development.Their diversity in structure and bioactivity,to a large extent,arise from oxidative modifications which are mainly catalyzed by cytochrome P450s(CYPs).The medicinal plant Euphorbia fischeriana Steud.is rich in LRDs with distinct scaffolds.Herein,we characterized three cytochrome P450s involved in LRD biosynthesis from this plant.Notably,CYP71D450 and CYP701A148 are two substrate-promiscuity CYPs.The former is the first example of CYPs which can oxidize C-3 of ent-atisane skeleton and ent-isopimara-7(8),15-diene,and the latter is the first example of CYPs which can oxidize C-19 of ent-abietane and ent-pimarane skeletons.This study expands the toolkit for bioproduction of diverse LRDs.
基金supported by grants from the Guangxi Natural Science Foundation(2024GXNSFAA010150)the Guangdong Basic and Applied Basic Research Foundation(2022A1515111167).
文摘Background:A significant proportion of patients still cannot benefit from existing targeted therapies and immunotherapies,making the search for new treatment strategies extremely urgent.In this study,we combined integrate public data analysis with experimental validation to identify novel prognostic biomarkers and therapeutic targets for lung adenocarcinoma(LUAD).Methods:We analyzed RNA and protein databases to assess the expression levels of cytochrome C oxidase 5B(COX5B)in LUAD.Several computational algorithms were employed to investigate the relationship between COX5B and immune infiltration in LUAD.To further elucidate the role of COX5B in LUAD,we utilized multiple experimental approaches,including quantitative reverse transcription PCR assays,western blot,immunohistochemistry,electron microscopy,flow cytometry,and EdU proliferation assays.Results:We revealed that COX5B was significantly elevated in LUAD and positively correlated with poor prognosis of LUAD patients.Analysis of co-expression network indicated that COX5B may take part in the intracellular adenosine triphosphate(ATP)synthesis through the oxidative phosphorylation pathway.There was a negative correlation between COX5B expression and immune infiltration in LUAD.Furthermore,we validated that COX5B levels were significantly elevated in both LUAD tissues and cell lines.Specifically,immunohistochemistry(IHC)assays revealed a 2.32-fold increase of COX5B in tumor tissues compared to that in adjacent normal tissues(p=0.0044).Additionally,COX5B knockdown disrupted the redox homeostasis,ultimately suppressed the proliferation of LUAD cells.Subsequent investigations demonstrated that berberine effectively targeted COX5B,diminishing its protein expression and consequently inhibiting cell proliferation and tumor growth in LUAD.Conclusions:This study established that upregulated COX5B was positive associated with poor patient prognosis in LUAD,elucidating the mechanisms by which berberine targets COX5B to inhibit tumor growth,thereby providing a novel therapeutic target and strategy for the clinical management of LUAD.
基金Supported by the Key R&D Program of Shandong Province(No.2023CXGC010411)the National Key R&D Program of China(Nos.2023YFD2400800,2022YFD2401301,2022FY100304)+2 种基金the National Natural Science Foundation of China(Nos.42076092,41906083,41776179)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDB42000000)the Earmarked Fund for Modern Agro-industry Technology Research System(No.CARS-47)。
文摘The Saccostrea mordax Gould,1850 is a typical intertidal species,whose genetic differentiation is influenced by various factors,including geological and climatic changes.To explore the genetic structure and historical population characteristics of Saccostrea mordax,we sequenced the mitochondrial cytochrome c oxidase subunit I(COI)gene from 58 specimens sampled from four locations in the western Pacific.Additionally,103 individuals from the Persian Gulf and western Pacific(from databases)were included for phylogenetic analysis.The Bayesian Inference tree showed that all specimens were divided into two clades,i.e.,the Persian Gulf population and the western Pacific population.Spatial molecular variance analysis indicated significant genetic differentiation between the two populations,and isolation by distance analysis revealed a positive correlation between genetic differentiation and geographic distance.Neutrality tests and Bayesian Skyline Plot suggested that both populations underwent expansions during the late Pleistocene.This study revealed the population history of Saccostrea mordax and described a new lineage,Saccostrea mordax lineage D,providing a foundation for understanding oyster biodiversity formation and genetic resource conservation.
基金supported by the National Natural Science Foundation of China(Grant Nos.81872831 and 82030107)the National Science and Technology Major Projects for‘significant new drugs creation’of the 13th five-year plan(2017ZX09101001 and 2018ZX09721002007,China)。
文摘D-a-Tocopheryl polyethylene glycol 1000 succinate(TPGS,also known as vitamin E-TPGS)is a biodegradable amphiphilic polymer prepared by esterification of vitamin E with polyethylene glycol(PEG)1000.It is approved by the US Food and Drug Administration(FDA)and has found wide application in nanocarrier drug delivery systems(NDDS).Fully characterizing the in vivo fate and pharmacokinetic behavior of TPGS is important to promote the further development of TPGS-based NDDS.However,to date,a bioassay for the simultaneous quantitation of TPGS and its metabolite,PEG1000,has not been reported.In the present study,we developed such an innovative bioassay and used it to investigate the pharmacokinetics,tissue distribution and excretion of TPGS and PEG1000 in rat after oral and intravenous dosing.In addition,we evaluated the interaction of TPGS with cytochromes P450(CYP450s)in human liver microsomes.The results show that TPGS is poorly absorbed after oral administration with very low bioavailability and that,after intravenous administration,TPGS and PEG1000 are mainly distributed to the spleen,liver,lung and kidney before both being slowly eliminated in urine and feces as PEG1000.In vitro studies show the inhibition of human CYP450 enzymes by TPGS is limited to a weak inhibition of CYP3A4.Overall,our results provide a clear picture of the in vivo fate of TPGS which will be useful in evaluating the safety of TPGS-based NDDS in clinical use and in promoting their further development.
基金This study was supported by the Army Research Office and was accomplished under grant number W911NF‐17‐1‐0345.
文摘Direct interspecies electron transfer(DIET)may be most important in methanogenic environments,but mechanistic studies of DIET to date have primarily focused on cocultures in which fumarate was the terminal electron acceptor.To better understand DIET with methanogens,the transcriptome of Geobacter metallireducens during DIET‐based growth with G.sulfurreducens reducing fumarate was compared with G.metallireducens grown in coculture with diverse Methanosarcina.The transcriptome of G.metallireducens cocultured with G.sulfurreducens was significantly different from those with Methanosarcina.Furthermore,the transcriptome of G.metallireducens grown with Methanosarcina barkeri,which lacks outer‐surface c‐type cytochromes,differed from those of G.metallireducens cocultured with M.acetivorans or M.subterranea,which have an outer‐surface c‐type cytochrome that serves as an electrical connect for DIET.Differences in G.metallireducens expression patterns for genes involved in extracellular electron transfer were particularly notable.Cocultures with c‐type cytochrome deletion mutant strains,ΔGmet_0930,ΔGmet_0557 andΔGmet_2896,never became established with G.sulfurreducens but adapted to grow with all three Methanosarcina.Two porin–cytochrome complexes,PccF and PccG,were important for DIET;however,PccG was more important for growth with Methanosarcina.Unlike cocultures with G.sulfurreducens and M.acetivorans,electrically conductive pili were not needed for growth with M.barkeri.Shewanella oneidensis,another electroactive microbe with abundant outer‐surface c‐type cytochromes,did not grow via DIET.The results demonstrate that the presence of outer‐surface c‐type cytochromes does not necessarily confer the capacity for DIET and emphasize the impact of the electron‐accepting partner on the physiology of the electron‐donating DIET partner.
文摘P450 enzymes-catalyzed aromatic hydroxylation plays an important role in detoxification,biosynthesis,and potential carcinogenic effect of aromatic compounds.Though it has been explored for decades,the actual process of aromatic hydroxylation and mechanism of regioselectivity catalyzed by cytochrome P450 monooxygenases remained ambiguous.Here,we have resolved these issues.With a stable chiral organofluorine probe,and especially with X-ray data of two isolated arene oxides derivatives,we demonstrate that an arene oxide pathway is definitely involved in P450-catalyzed aromatic hydroxylation.By the capture,isolation,identification and reactivity exploration of the arene 1,2-oxide and arene 2,3-oxide intermediates,together with advanced QM calculations,the mechanism of how two intermediates go to the same product has been elucidated.In addition to the model substrate,we also confirmed that an arene oxide intermediate is involved in the P450-catalyzed hydroxylation pathway of a natural product derivative methyl cinnamate,which indicates that this intermediate appears to be universal in P450-catalyzed aromatic hydroxylation.Our work not only provides the most direct evidence for the arene oxide pathway and new insights into the regioselectivity involved in P450-catalyzed aromatic hydroxylation,but also supplies a new synthetic approach to achieve the dearomatization of aromatic compounds.
基金supported by the Sichuan Province International Science and Technology Innovation Cooperation(2024YFHZ0299)the Project of Science and Technology Department of Sichuan Province(2022YFH0031)Chengdu Science and Technology Bureau(2024-YF05-02168-SN).
文摘Rice sheath blight(RSB)is a major destructive disease impeding rice production.Identifying key germplasm resources with increased resistance remains a challenge.However,the mechanisms underlying disease resistance are not yet fully understood.Cytochrome P450 monooxygenases(CYP450s)serve biosynthesis and metabolic detoxification functions in plants,but there is limited information about their role in the response induced by RSB.This study demonstrated that CYT02 belongs to the CYP73A100 subfamily and is a typical member of the CYP450s.Overexpression(OE)in rice of the cytochrome P450 monooxygenase cyt02 conferred increased resistance to RSB and increased vegetative tillering.Cyt02 may increase RSB resistance by regulating plant hormone synthesis,regulate reactive oxygen species(ROS)by coordinating the activity of antioxidant enzymes,and initiate phytoalexin synthesis in response to fungal infection.These research findings have laid a foundation for a deeper understanding of the function of cyt02 and offered a potential target gene for breeding rice varieties resistant to sheath blight.
文摘Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Iα-loop-helix Iβmotif which corresponds to helix I of other P 450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,theγ-domain.The structure reveals a homodimer in which theγ-domain of one molecule interacts with theβ-domain of another.The plane of heme group makes an angle of approximately 40°with the helix Iα-loop-helix Iβmotif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recognition.Phe-301,Ser-303 and Gly-305 from the helix Iα-loop-helix Iβmotif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.
基金supported by the National Natural Science Foundation of China(No.81872584)the Natural Science Foundation of Shenzhen(No.JCYJ20210324093211030)+1 种基金Henan Province’s Key R&D and Promotion Projects(No.252102310075)the Interdisciplinary Research for Firstclass Discipline Construction Project of Henan University(No.2019YLXKJC04)。
文摘A class of persistent organic pollutants,polychlorinated biphenyls(PCBs),are ubiquitous in the environment and human tissues which are continuously and long-term threatened.We aim to investigate the toxic effects and the underlying mechanisms of PCB153 using Caenorhabditis elegans as a model organism.Our findings demonstrated that exposure to an optimized concentration of 2μmol/L PCB153 had adverse effects on C.elegans,led to reduction of lifespan,body length,frequency of body bending,and head wiggling.Additionally,this exposure led to an increase in the accumulation of reactive oxygen species,superoxide dismutase,lipofuscin and fat content within the organism.Furthermore,gene set enrichment analysis and whole transcriptome sequencing revealed,a significant up-regulation of Cytochrome P450(CYP)family genes,which are crucial for the xenobiotics metabolism in C.elegans.Knocking down specific genes within the CYPs family,including cyp-35C1,cyp-35A5,cyp-35A3,cyp-34A1,and cyp-34A10,via RNA interference were able to reverse the shortening of lifespan and fat accumulation induced by PCB153 exposure.Moreover,a comprehensive a competing endogenous RNA network was constructed by integrating qPCR-validated differentially expressed m RNA along with lncRNA,circRNA,and miRNA identified by transcriptomic sequencing.This study emphasizes that PCB153 exposure leads to the physiological impairments,particularly fat accumulation and a shorter lifespan,through the modulation of CYP450 family genes.These findings contribute to our understanding of environmental pollutants and their impact on biological systems and provide valuable information on the toxic effects and potential risks associated with PCB153 exposure.
基金supported by the Rufford Foundation(Grant No.45888-2:data analysis)the Russian Science Foundation(RSF Grant No.22-14-00037-P:data analysis)+3 种基金the National Natural Science Foundation of China(32300370)International Partnership Program of Chinese Academy of Sciences(071GJHZ2023041MI),ChinaPostdoctoral Science Foundation(2023M743416)the 2023 award fund of Qinghai Provincial Key Laboratory of Animal Ecological Genomics.
文摘A comprehensive taxonomic reassessment of the Hebius optatus species complex,integrating molecular data and morphological data,has led to the recognition and description of a previously overlooked species from northern Vietnam and the Guangxi Zhuang Autonomous Region,China,herein described as Hebius vogeli sp.nov.Phylogenetic analyses reveal that Hebius vogeli sp.nov.is closely related to H.sauteri and H.optatus sensu stricto.Uncorrected p-distances based on cytochrome b sequences indicate a divergence of 10.74%-11.73%between Hebius vogeli sp.nov.and H.optatus.The new species differs from H.optatus by a combination of characters,including a higher number of subcaudal scales,a greater total number of ventral and subcaudal scales,and distinct ventral coloration.Hebius vogeli sp.nov.appears to be geographically restricted to northern Vietnam and the adjacent region of Guangxi Zhuang Autonomous Region,China,making its distribution allopatric from that of H.optatus.The Pearl River is hypothesized to serve as a biogeographic barrier separating the two taxa.Accordingly,we recommend the exclusion of H.optatus from the herpetofauna of Vietnam.Based on IUCN Red List criteria,we propose that Hebius vogeli sp.nov.be assessed as Least Concern(LC).Our results emphasize both the underestimated diversity within Hebius and the need for continued integrative taxonomic efforts,particularly in the Vietnam-China borderlands,where cryptic diversity remains largely unexplored.
基金Supported by the Natural Science Foundation of Hebei Province Beijing-Tianjin-Hebei Basic Research Special Program,No.H2020110287Key Laboratory for Diabetic Kidney Disease Syndrome and Treatment of the Traditional Chinese Medicine Administration of Hebei Province,No.7[2024]+2 种基金Yuansong Wang National Famous Traditional Chinese Medicine Expert Heritage Studio,No.3[2024]Hebei Key Laboratory of Integrated Traditional Chinese and Western Medicine for Diabetes and Its Complications,No.SZX2020015Jiangsu Province Chinese Medicine Science and Technology Development Program Project,No.YB2020065.
文摘BACKGROUND Diabetic nephropathy(DN)is a major complication of diabetes,marked by progressive renal damage and an inflammatory response.Although research has investigated the pathological mechanisms underlying DN,effective treatment options remain limited.AIM To evaluate the therapeutic impact of Shenzhuo formulation(SZF)on a DN mouse model and to examine its potential molecular mechanisms using transcriptomic and metabolomic approaches.METHODS We established a DN mouse model through a high-fat diet combined with streptozotocin(STZ)injection,followed by SZF treatment.We analyzed SZF’s effects on gene expression and metabolite profiles in renal tissues of DN mice using transcriptomics and metabolomics techniques.Additionally,based on transcriptomic and non-targeted metabolomic findings,we further assessed SZF’s influence on the expression of factors related to the cytochrome P450(CYP450)-mediated arachidonic acid(AA)metabolism pathway,as well as its effects on inflammation and oxidative stress.RESULTS SZF intervention significantly decreased hyperglycemia and mitigated renal function impairment in DN mice.Pathological analysis revealed that SZF treatment improved renal tissue damage,reduced fibrosis,and diminished glycogen deposition.Transcriptomic analysis indicated that SZF influenced mRNA expression of CYP450-related genes,including Cyp2j13,Cyp2b9,Pla2 g2e/Cyp4a12a,Cyp4a32,Cyp2e1,and Cyp4a14.Non-targeted metabolomic results demonstrated that SZF altered the levels of metabolites associated with the AA metabolic pathway,including 5,6-EET,14,15-EET,phosphatidylcholine,and 20-HETE.Further experiments showed that SZF upregulated the expression of CYP4A and CYP2E proteins in renal tissue,as well as CYP2J and CYP2B proteins.Additionally,SZF significantly reduced the expression of inflammatory factors in renal tissue,enhanced antioxidant enzyme activity,and alleviated oxidative stress.CONCLUSION SZF exerts anti-inflammatory and antioxidant effects by regulating CYP450-mediated AA metabolism,leading to improved renal function and improved pathological state in DN mice.
文摘The cytochrome P4503A(CYP3A)gene family’s role in early progression of gastric cancer was comprehensively investigated.Its potential as a therapeutic target was evaluated.Upon literature review,aberrant expression of the CYP3A gene family has a strong correlation with gastric cancer onset,although the precise underlying mechanisms remain unclear.To assess its potential as a biomarker for early diagnosis and a therapeutic target,we have provided a comprehensive review of the regulatory mechanisms governing CYP3A gene family expression in gastric cancer,as well as its relation with early tumor progression and the tumor microenvironment.The CYP3A gene family is crucial in the proliferation,migration,and invasion of gastric cancer cells and promotes cancer progression by modulating inflammatory responses and oxidative stress within the tumor microenvironment.Furthermore,genetic polymorphisms in CYP3A enzymes highlight its potential value in personalized medicine.Based on these findings,this paper explores the feasibility of developing inhibitors and activators targeting CYP3A enzymes and discusses potential applications in gene therapy.This research provides crucial theoretical support for the CYP3A gene family as an early diagnostic marker and therapeutic target for gastric cancer.In the future,multi-omics studies and large-scale clinical trials will be essential to advance clinical translation of these findings.
文摘Heme peroxygenases exhibit remarkable catalytic versatility in facilitating a wide array of oxidative reactions under mild conditions,eliminating the need for coenzymes and intricate electron transport systems.This unique character underscores their essentiality and potential as promising tools in synthetic biology.Recent advancements in enzyme engineering have significantly enhanced the catalytic performance of both natural and artificial peroxygenases.Extensive engineering efforts have been directed towards unspecific peroxygenases and fatty acid peroxygenases,aiming to expand their substrate specificities,and enhance reaction selectivities,as well as increase enzyme stability.Furthermore,innovative strategies such as dual-functional small molecule-assisted systems and H_(2)O_(2) tunnel engineering have been harnessed to transform P450 monooxygenases into highly efficient peroxygenases,capable of catalyzing reactions with a variety of unnatural substrates.This review consolidates the latest progress in the engineered and artificial heme peroxygenases,emphasizing their catalytic performances as potent biocatalysts for sustainable organic synthesis.
基金partly supported by research grants from the Outstanding Youth Foundation of the Beijing Academy of Agriculture and Forestry Sciences,China(YXQN202301)the National Natural Science Foundation of China(32272522)the Beijing Natural Science Foundation,China(6232005)。
文摘The tobacco whitefly,Bemisia tabaci,is a notorious pest affecting various crops globally,and it exhibits high levels of resistance to various insecticides.Afidopyropen is a recently commercialized pyropene insecticide for B.tabaci control with high selectivity and a novel mode of action.We previously identified a high level of afidopyropen resistance in a field-collected population after selection in the lab,and named it the HD-Afi strain.In the present study,minimal cross-resistance in the HD-Afi strain was found between afidopyropen and other common chemical agents.However,the P450 enzyme activity in HD-Afi was 2.18 times the level in susceptible strain HD-S.Expression analysis revealed that two of 12 candidate P450 genes,namely CYP6DW3 and CYP4C64,were significantly up-regulated in HD-Afi.Silencing CYP6DW3 and CYP4C64 by RNA interference(RNAi)substantially increased the susceptibility of whitefly adults,confirming their involvement in afidopyropen resistance.Homology modeling and molecular docking analyses demonstrated stable binding of afidopyropen to CYP6DW3 and CYP4C64,with binding free energies of–6.87 and–6.11 kcal mol^(-1),respectively.The findings of this study suggest that the induction of CYP6DW3 and CYP4C64 facilitates afidopyropen detoxification,contributing to the development of resistance in B.tabaci.
基金supported by the National Natural Scientific Foundation of China(32160254)Yunnan Fundamental Research Projects(202401AS070039).
文摘Background:The thermogenic characteristics of animals are closely related to species distribution,and basal metabolic rate(BMR)and thermal neutral zone(TNZ)are important components of the thermogenic process in animals.Furthermore,the cytochrome c oxidase 1(COX1)gene has become a subject of particular interest due to its high degree of sequence conservation,stable evolutionary rate,and rare insertions/deletions.Method:The present study selected 29 species of Rodentia and 20 species of Chiroptera.The present study employed the statistical software SPSS(27.0 Chinese version)and Origin(2024 Chinese version)software to conduct correlation analyses on a variety of biological and ecological variables.These variables included body weight,BMR,TNZ,upper thermal neutral zone,lower thermal neutral zone(LTNZ),litter size,and dietary patterns of species from different latitudes.Furthermore,we conducted a series of phylogenetic tree analyses on COX1 protein.Results:As the geographic location of a species increases in latitude,there is an observed upward trend in both body mass and BMR of rodent species.Rodentia and Chiroptera species have been observed to exhibit a decrease in LTNZ and an expansion of TNZ.With regard to dietary habits,Rodentia species are predominantly phytophagous or omnivorous.Omnivorous species exhibit a marked tendency to produce larger litters in comparison to their herbivorous counterparts.Chiroptera species exhibit a diverse dietary range,including phytophagous,carnivorous,and omnivorous species.However,no correlation was observed between dietary differences and litter size.Phylogenetic analysis of the COX1 protein subsequently demonstrated that these two species groups share a monophyletic origin.Conclusion:The present study suggests that the selected Rodentia and Chiroptera species adapt to high-latitude environments by lowering the LTNZ and widening the TNZ.Furthermore,an upward trend has been observed in the body mass and BMR of high-latitude Rodentia.Phylogenetic analysis of the COX1 protein across various taxonomic groups substantiates the efficacy of this gene for species identification.Integrating physiological phenotypes with COX1 protein molecular evidence,this study provides a reference framework for the multidimensional mechanisms of mammalian latitudinal adaptation.
基金supported by grants from the National Natural Science Foundation of China(32460505)China Agriculture Research System(CARS-11)+2 种基金the Chinese Academy of Tropical Agricultural Sciences for the Science and Technology Innovation Team of the National Tropical Agricultural Science Center(CATASCXTD202301)Additional support was provided by the Hainan Province Graduate Innovation Research Project(Hyb2020-09)the Key Laboratory of Tropical Fruits and Vegetables Quality and Safety for State Market Regulation(KF-2023016).
文摘The high content of cyanogenic glycosides(CG)in cassava tubers affects food safety.CG are involved in the plant growth and development and protect cassava leaves from herbivorous predators.However,the regulatory mechanism of CG biosynthesis remains poorly understood.Here,yeast one-hybrid assays were performed using a mixed cDNA library of cassava tubers and leaves as prey and the promoter of MeCYP79D2 as bait.MeCYP79D2,a cytochrome P450 protein,is the rate-limiting enzyme for CG synthesis in cassava.From this information,a candidate regulator of MeCYP79D2 was selected and identified as transcription factor MePHD1.2.MePHD1.2,located in the nucleus and exhibiting an inhibitory transcription activity directly bound to an AT-rich motif in the promoter of MeCYP79D2.In cassava,the transcriptional activity of MeCYP79D2 was considerably enhanced in mephd1.2 mutant lines leading to increased linamarin and lotaustralin contents.Deletion of MePHD1.2 promoted the production of CGs in cassava and decreased transcription inhibition on MeCYP79D2,exposing a novel regulatory module governing biosynthesis of CGs.
文摘Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches.CYP80B,a family of cytochrome P450(CYP450)enzymes,plays a crucial role in BIA biosynthesis.Previously reported CYP80Bs are known to catalyze the 3′-hydroxylation of(S)-Nmethylcoclaurine,with the N-methyl group essential for catalytic activity.In this study,we successfully cloned a full-length CYP80B gene(St CYP80B)from Stephania tetrandra(S.tetrandra)and identified its function using a yeast heterologous expression system.Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that St CYP80B could catalyze Nmethylcoclaurine and coclaurine into their respective 3'-hydroxylated products.Notably,St CYP80B exhibited an expanded substrate selectivity compared to previously reported wildtype CYP80Bs,as it did not require an N-methyl group for hydroxylase activity.Furthermore,St CYP80B displayed a clear preference for the(S)-configuration.Co-expression of St CYP80B with the CYP450 reductases(CPRs,StCPR1,and StCPR2),also cloned from S.tetrandra,significantly enhanced the catalytic activity towards(S)-coclaurine.Site-directed mutagenesis of St CYP80B revealed that the residue H205 is crucial for coclaurine catalysis.Additionally,St CYP80B exhibited tissue-specific expression in plants.This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
