NLRP3 inflammasome activation is pivotal for cytokine secretion and pyroptosis in response to diverse stimuli,playing a crucial role in innate immunity.While extensively studied in mammals,the regulatory mechanisms go...NLRP3 inflammasome activation is pivotal for cytokine secretion and pyroptosis in response to diverse stimuli,playing a crucial role in innate immunity.While extensively studied in mammals,the regulatory mechanisms governing NLRP3 activation in non-mammalian vertebrates remain largely unexplored.Teleosts,as basal vertebrates,represent an ideal model for exploring the evolutionary trajectory of inflammasome regulation.In this study,ABE assays,confocal microscopy,and biochemical analyses were applied to systematically characterize the mechanisms underlying NLRP3 inflammasome in teleosts,using large yellow croakers(Larimichthys crocea,Lc)and zebrafish(Danio rerio,Dr)as representative models.Our findings revealed a previously unrecognized palmitoylation-dependent regulatory mechanism essential for teleost NLRP3 activation.Specifically,zDHHC18-mediated palmitoylation at a teleost-specific cysteine residue(C946 in LcNLRP3,C1037 in DrNLRP3)was required for the translocation of NLRP3 to the dispersed trans-Golgi network,facilitating its subsequent recruitment to the microtubule-organizing center.This membrane trafficking was crucial for inflammasome assembly and downstream inflammatory responses.These findings provide new insights into the distinct regulatory mechanisms of NLRP3 activation in teleosts,highlighting an evolutionary divergence that contributes to innate immunity adaptation in early vertebrates.展开更多
The partitioning of membrane proteins into lipid domains in cellular membranes is closely associated with the realization of the protein functions and it is influenced by various factors such as the post-translational...The partitioning of membrane proteins into lipid domains in cellular membranes is closely associated with the realization of the protein functions and it is influenced by various factors such as the post-translational modification of palmitoylation.However,the molecular mechanism of the effect of palmitoylation on membrane protein partitioning into the lipid domains remains elusive.In this work,taking human peripheral myelin protein 22(PMP22)as an example,we employ coarse-grained molecular dynamics simulations to investigate the partitioning of both the natural PMP22 and the palmitoylated PMP22(pal-PMP22)into the lipid domains of model myelin membranes.The results indicate that palmitoylation drives PMP22 to localize at the boundary of the liquid-ordered(Lo)and liquid-disordered(Ld)domains and increases the possibility of PMP22 partitioning into the Lo domains by changing the hydrophobic length of the proteins and perturbing the ordered packing of tails of the saturated lipids in the Lo domains.This work offers some novel insights into the role of palmitoylation in modulating the function of membrane proteins in cellular membranes.展开更多
Multicellular organisms use chemical messengers to transmit signals among organelles and to other cells. Relatively small hydrophobic molecules such as lipids are excellent candidates for this signaling purpose. In mo...Multicellular organisms use chemical messengers to transmit signals among organelles and to other cells. Relatively small hydrophobic molecules such as lipids are excellent candidates for this signaling purpose. In most proteins, palmitic acid and other saturated and some unsaturated fatty acids are esterified to the free thiol of cysteines and to the N-amide terminal. This palmitoylation process enhances the surface hydrophobicity and membrane affinity of protein substrates and plays important roles in modulating proteins’ trafficking, stability, and sorting etc. Protein palmitoylation has been involved in numerous cellular processes, including signaling, apoptosis, and neuronal transmission. The palmitoylation process is involved in multiple diseases such as Huntington’s disease, various cardiovascular and T-cell mediated immune disorders, as well as cancer. Protein palmitoylation through the thioester (S-acylation) is unique in that it is the only reversible lipid modification. Our study on lipopolysaccharide (LPS) and deoxynivalenol (DON) treatment to rats provides some insights to the complex role of protein palmitoylation in chemical and microbial toxicity. In contrast, myrisoylated proteins contain the 14-carbon fatty acid myristate attached via amide linkage to the N-terminal glycine residue of protein, and occur cotranslationally. The bacterial outer membrane enzyme lipid A palmitoyltransferase (PagP) confers resistance to host immune defenses by transferring a palmitate chain from a phospholipid to the lipid A component of LPS. PagP is sensitive to cationic antimicrobial peptides (CAMP) which are included among the products of the Toll-like receptor 4 (TLR4) signal transduction pathway. This modification of lipid A with a palmitate appears to both and protects the pathogenic bacteria from host immune defenses and attenuates the activation of those same defenses through the TLR4 signal transduction pathway.展开更多
Palmitoylation plays a key role in liver cancer(HCC)initiation,progression,and drug resistance.This review highlights palmitoyltransferases(PATs),like ZDHHC family enzymes,and their impact on HCC and immune evasion.We...Palmitoylation plays a key role in liver cancer(HCC)initiation,progression,and drug resistance.This review highlights palmitoyltransferases(PATs),like ZDHHC family enzymes,and their impact on HCC and immune evasion.We discuss small-molecule inhibitors targeting these enzymes,such as ZDHHC9 and PPT1 inhibitors,showing promising results.The role of palmitoylation in immune checkpoint molecules like TIM-3 is explored,along with its potential in combination therapy.Additionally,we examine palmitoylation-related biomarkers for non-invasive diagnosis,prognosis,and treatment monitoring in liver cancer.展开更多
Palmitoylation,an essential covalent attachment of a fatty acid(usually C16 palmitate)to cysteine residues within proteins,is crucial for regulating protein functionality and enzymatic activities.This lipid modificati...Palmitoylation,an essential covalent attachment of a fatty acid(usually C16 palmitate)to cysteine residues within proteins,is crucial for regulating protein functionality and enzymatic activities.This lipid modification facilitates the anchoring of proteins to cellular membranes,dictating their subcellular distribution and influencing protein transport dynamics and intracellular positioning.Additionally,it plays a role in regulating protein degradation through the ubiquitin-proteasome system.Palmitoylation is implicated in the pathogenesis and progression of cardiovascular diseases by modulating substrates and prompting additional post-translational modifications,as well as by interacting with other molecular alterations.Moreover,an intervention strategy focusing on palmitoylation processes is anticipated to offer novel therapeutic avenues for cardiovascular pathologies and address extant challenges in clinical settings.This review consolidates current research on the role and importance of palmitoylation in cardiovascular diseases by exploring its regulatory functions,the catalyzing enzymes,and the involved substrates.It highlights recent discoveries connecting palmitoylation-targeted therapies to cardiovascular health and examines potential approaches and future challenges in cardiovascular treatment.展开更多
Fatty acid metabolism mediates macrophage function;however,the underlying mechanism by which fatty acid metabolism regulates macrophage interleukin(IL)-1βproduction remains to be uncovered.Here,we used genome-wide as...Fatty acid metabolism mediates macrophage function;however,the underlying mechanism by which fatty acid metabolism regulates macrophage interleukin(IL)-1βproduction remains to be uncovered.Here,we used genome-wide association studies(GWAS)to identify several porcine serum IL-1β-related genes,such as the fatty acid metabolizing enzyme acyl-CoA thioesterase 11(ACOT11).We then demonstrated that inflammatory macrophages have low expression of ACOT11,and ACOT11 overexpression inhibits IL-1βmaturation from inflammatory macrophages.Mechanistically,ACOT11 promotes intracellular fatty acids accumulation,including eicosatetraenoic acid(EA)and stearic acid(SA),which inhibit activation of the Janus kinase(JAK)—signal transducer and activator of transcription(STAT)signaling through palmitoylation of interferon(IFN)-γreceptor(IFNGR)2 at C261site.Furthermore,we also found that EA attenuates lipopolysaccharide(LPS)-induced sepsis in mice.Collectively,our findings reveal a mechanism involving ACOT11-mediated post-translational modification that regulates macrophage function and provide a promising therapeutic target for the treatment of inflammatory diseases associated with macrophages.展开更多
Nutrients from dietary foods not only provide energy and building blocks,but also play critical roles in modulating diverse pathophysiological functions.They achieve these,in part,by accelerating cell signaling transd...Nutrients from dietary foods not only provide energy and building blocks,but also play critical roles in modulating diverse pathophysiological functions.They achieve these,in part,by accelerating cell signaling transduction processes via modulating various types of protein post-translational modifications(PTMs).Notably,accumulating evidence has identified palmitic acid(PA),a major component of high-fat diets,as a significant contributor to various human disorders,including diabetes and cancer.Hence,further understanding the roles of PA and its involvement in protein palmitoylation,a key PTM,is crucial for uncovering the mechanisms underlying these diseases and exploring potential clinical applications in cancer therapy.This review comprehensively summarizes recent advances in the understanding of PA homeostasis and palmitoylation in tumorigenesis.Specifically,it highlights the connections between palmitoylation and key processes such as oncogenic signaling pathways,cell death mechanisms,innate immune responses,and the tumor microenvironment.The review also emphasizes potential therapeutic strategies,including targeting PA homeostasis,palmitoylation-associated processes,or specific palmitoylated proteins for cancer treatment.Finally,the challenges in the field,such as the regulation of PA homeostasis and the dynamic detection or targeting of palmitoylation,are discussed,underscoring the need for further research to address these critical issues.展开更多
Protein palmitoylation is a widespread lipid modification in which one or more cysteine thiols on a substrate protein are modified to form a thioester with a palmitoyl group.This lipid modification is readily reversib...Protein palmitoylation is a widespread lipid modification in which one or more cysteine thiols on a substrate protein are modified to form a thioester with a palmitoyl group.This lipid modification is readily reversible;a feature of protein palmitoylation that allows for rapid regulation of the function of many cellular proteins.Mutations in palmitoyltransferases(PATs),the enzymes that catalyze the formation of this modification,are associated with a number of neurological diseases and cancer progression.This review summarizes the crucial role of palmitoylation in biological systems,the discovery of the DHHC protein family that catalyzes protein palmitoylation,and the development of methods for investigating the catalytic mechanism of PATs.展开更多
Epithelial ovarian cancer(EOC) exhibits strong dependency on the tricarboxylic acid(TCA) cycle and oxidative phosphorylation to fuel anabolic process.Here,we show that malate dehydrogenase 2(MDH2),a key enzyme of the ...Epithelial ovarian cancer(EOC) exhibits strong dependency on the tricarboxylic acid(TCA) cycle and oxidative phosphorylation to fuel anabolic process.Here,we show that malate dehydrogenase 2(MDH2),a key enzyme of the TCA cycle,is palmitoylated at cysteine 138(C138) residue,resulting in increased activity of MDH2.We next identify that ZDHHC18 acts as a palmitoyltransferase of MDH2.Glutamine deprivation enhances MDH2 palmitoylation by increasing the binding between ZDHHC18 and MDH2.MDH2 silencing represses mitochondrial respiration as well as ovarian cancer cell proliferation both in vitro and in vivo.Intriguingly,re-expression of wild-type MDH2,but not its palmitoylation-deficient C138 S mutant,sustains mitochondrial respiration and restores the growth as well as clonogenic capability of ovarian cancer cells.Notably,MDH2 palmitoylation level is elevated in clinical cancer samples from patients with high-grade serous ovarian cancer.These observations suggest that MDH2 palmitoylation catalyzed by ZDHHC18 sustains mitochondrial respiration and promotes the malignancy of ovarian cancer,yielding possibilities of targeting ZDHHC18-mediated MDH2 palmitoylation in the treatment of EOC.展开更多
Glutamate acting on AMPA-type ionotropic glutamate receptor(AMPAR) mediates the majority of fast excitatory synaptic transmission in the mammalian central nervous system. Dynamic regulation of AMPAR by post-translatio...Glutamate acting on AMPA-type ionotropic glutamate receptor(AMPAR) mediates the majority of fast excitatory synaptic transmission in the mammalian central nervous system. Dynamic regulation of AMPAR by post-translational modifications is one of the key elements that allow the nervous system to adapt to environment stimulations. S-palmitoylation, an important lipid modification by post-translational addition of a long-chain fatty acid to a cysteine residue, regulates AMPA receptor trafficking, which dynamically affects multiple fundamental brain functions, such as learning and memory. In vivo, S-palmitoylation is controlled by palmitoyl acyl transferases and palmitoyl thioesterases.In this review, we highlight advances in the mechanisms for dynamic AMPA receptors palmitoylation,and discuss how palmitoylation affects AMPA receptors function at synapses in recent years.Pharmacological regulation of S-palmitoylation may serve as a novel therapeutic strategy for neurobiological diseases.展开更多
Gain-of-function mutations in the STING-encoding gene TMEM173 are central to the pathology of the autoinflammatory disorder STING-associated vasculopathy with onset in infancy(SAVI).Furthermore,excessive activity of t...Gain-of-function mutations in the STING-encoding gene TMEM173 are central to the pathology of the autoinflammatory disorder STING-associated vasculopathy with onset in infancy(SAVI).Furthermore,excessive activity of the STING signaling pathway is associated with autoinflammatory diseases,including systemic lupus erythematosus and Aicardi–Goutières syndrome(AGS).Two independent studies recently identified pharmacological inhibitors of STING.Strikingly,both types of compounds are reactive nitrocontaining electrophiles that target STING palmitoylation,a posttranslational modification necessary for STING signaling.As a consequence,the activation of downstream signaling molecules and the induction of type I interferons were inhibited.The compounds were effective at ameliorating inflammation in a mouse model of AGS and in blocking the production of type I interferons in primary fibroblasts from SAVI patients.This mini-review focuses on the roles of palmitoylation in STING activation and signaling and as a pharmaceutical target for drug development.展开更多
The TEA domain(TEAD)family proteins(TEAD1-4)are essential transcription factors that control cell differentiation and organ size in the Hippo pathway.Although the sequences and structures of TEAD family proteins are h...The TEA domain(TEAD)family proteins(TEAD1-4)are essential transcription factors that control cell differentiation and organ size in the Hippo pathway.Although the sequences and structures of TEAD family proteins are highly conserved,each TEAD isoform has unique physiological and pathological functions.Therefore,the development and discovery of subtype selective inhibitors for TEAD protein will provide important chemical probes for the TEAD-related function studies in development and diseases.Here,we identified a novel TEAD 1/3 covalent inhibitor(DC-TEADin1072)with biochemical IC50 values of 0.61±0.02 and 0.58±0.12μmol/L against TEAD1 and TEAD3,respectively.Further chemical optimization based on DC-TEAD in 1072 yielded a selective TEAD3 inhibitor DCTEAD3 in03 with the IC_(50) value of 0.16±0.03μmol/L,which shows 100-fold selectivity over other TEAD isoforms in activity-based protein profiling(ABPP)assays.In cells,DC-TEAD3 in03 showed selective inhibitory effect on TEAD3 in GAL4-TEAD(1-4)reporter assays with the IC50 value of1.15μmol/L.When administered to zebrafish juveniles,experiments showed that DC-TEAD3 in03 reduced the growth rate of zebrafish caudal fins,indicating the importance of TEAD3 activity in controlling proportional growth of vertebrate appendages.展开更多
The native Goα was purified from bovine brain cortex and palmitoylated in vitro. The in vitro palmitoylation site was the same as that in vivo. The internal palmitoylation of purified native Goα was found to be larg...The native Goα was purified from bovine brain cortex and palmitoylated in vitro. The in vitro palmitoylation site was the same as that in vivo. The internal palmitoylation of purified native Goα was found to be largely maintained. The apparent palmitoylation ratio was significantly increased after the Goα was treated with DTT. The GTPγS binding characteristic of Goα was not influenced by palmitoylation, however, the affinity for LUVs was increased dramatically. The in vitro palmitoylation model of Goα provides a better basis for studying the functional role of G protein palmitoylation in signal transduction.展开更多
Acyl-CoA synthetase long chain family member 5(ACSL5),is a member of the acyl-CoA synthetases(ACSs)family that activates long chain fatty acids by catalyzing the synthesis of fatty acyl-CoAs.The dysregulation of ACSL5...Acyl-CoA synthetase long chain family member 5(ACSL5),is a member of the acyl-CoA synthetases(ACSs)family that activates long chain fatty acids by catalyzing the synthesis of fatty acyl-CoAs.The dysregulation of ACSL5 has been reported in some cancers,such as glioma and colon cancers.However,little is known about the role of ACSL5 in acute myeloid leukemia(AML).We found that the expression of ACSL5 was higher in bone marrow cells from AML patients compared with that from healthy donors.ACSL5 level could serve as an independent prognostic predictor of the overall survival of AML patients.In AML cells,the ACSL5 knockdown inhibited cell growth both in vitro and in vivo.Mechanistically,the knockdown of ACSL5 suppressed the activation of the Wnt/β-catenin pathway by suppressing the palmitoylation modification of Wnt3a.Additionally,triacsin c,a pan-ACS family inhibitor,inhibited cell growth and robustly induced cell apoptosis when combined with ABT-199,the FDA approved BCL-2 inhibitor for AML therapy.Our results indicate that ACSL5 is a potential prognosis marker for AML and a promising pharmacological target for the treatment of molecularly stratified AML.展开更多
More than 85%of patients with uveal melanoma(UM)carry a GNAQ or GNA11 mutation at a hotspot codon(Q209)that encodes G proteinαsubunit q/11 polypeptides(Gα_(q/11)).GNAQ/11 relies on palmitoylation for membrane associ...More than 85%of patients with uveal melanoma(UM)carry a GNAQ or GNA11 mutation at a hotspot codon(Q209)that encodes G proteinαsubunit q/11 polypeptides(Gα_(q/11)).GNAQ/11 relies on palmitoylation for membrane association and signal transduction.Despite the palmitoylation of GNAQ/11 was discovered long before,its implication in UM remains unclear.Here,results of palmitoylation-targeted mutagenesis and chemical interference approaches revealed that the loss of GNAQ/11 palmitoylation substantially affected tumor cell proliferation and survival in UM cells.Palmitoylation inhibition through the mutation of palmitoylation sites suppressed GNAQ/11^(Q209L)-induced malignant transformation in NIH3T3 cells.Importantly,the palmitoylation-deficient oncogenic GNAQ/11 failed to rescue the cell death initiated by the knock down of endogenous GNAQ/11 oncogenes in UM cells,which are much more dependent on Gα_(q/11) signaling for cell survival and proliferation than other melanoma cells without GNAQ/11 mutations.Furthermore,the palmitoylation inhibitor,2-bromopalmitate,also specifically disrupted Gα_(q/11) downstream signaling by interfering with the MAPK pathway and BCL2 survival pathway in GNAQ/11-mutant UM cells and showed a notable synergistic effect when applied in combination with the BCL2 inhibitor,ABT-199,in vitro.The findings validate that GNAQ/11 palmitoylation plays a critical role in UM and may serve as a promising therapeutic target for GNAQ/11-driven UM.展开更多
Membrane-initiated estrogen receptorα(mERα)signaling has been shown to affect bone mass in murine models.However,it remains unknown which cell types mediate the mERα-dependent effects on bone.In this study,we gener...Membrane-initiated estrogen receptorα(mERα)signaling has been shown to affect bone mass in murine models.However,it remains unknown which cell types mediate the mERα-dependent effects on bone.In this study,we generated a novel mouse model with a conditional C451A mutation in Esr1,which enables selective knockout of the palmitoylation site essential for the membrane localization of ERα(C451A^(f/f)).First,we used Runx2-Cre mice to generate Runx2-C451A^(f/f)mice with conditional inactivation of mERαsignaling in Runx2-expressing osteoblast lineage cells.No significant changes were observed in body weight,weights of estrogen-responsive organs,or serum concentrations of estradiol between female Runx2-C451A^(f/f)and homozygous C451A^(f/f)littermate controls.High-resolution microcomputed tomography analysis showed a consistent decrease in cortical bone mass in the tibia,femur,and vertebra L5 of Runx2-C451A^(f/f)mice and three-point bending analysis of humerus revealed an impaired mechanical bone strength in Runx2-C451A^(f/f)female mice compared to controls.Additionally,primary osteoblast cultures from mice lacking mERαsignaling showed impaired differentiation compared to controls.展开更多
The host intestinal microbiota has emerged as the third element in the interactions between hosts and enteric viruses,and potentially affects the infection processes of enteric viruses.However,the interaction of porci...The host intestinal microbiota has emerged as the third element in the interactions between hosts and enteric viruses,and potentially affects the infection processes of enteric viruses.However,the interaction of porcine enteric coronavirus with intestinal microorganisms during infection remains unclear.In this study,we used 16S-rRNA-based Illumina NovaSeq high-throughput sequencing to identify the changes in the intestinal microbiota of piglets mediated by porcine epidemic diarrhea virus(PEDV)infection and the effects of the alterations in intestinal bacteria on PEDV infection and its molecular mechanisms.The intestinal microbiota of PEDV-infected piglets had significantly less diversity than the healthy group and different bacterial community characteristics.Among the altered intestinal bacteria,the relative abundance of Clostridium perfringens was significantly increased in the PEDV-infected group.A strain of C.perfringens type A,named DQ21,was successfully isolated from the intestines of healthy piglets.The metabolites of swine C.perfringens type A strain DQ21 significantly enhanced PEDV replication in porcine intestinal epithelial cell clone J2(IPEC-J2)cells,and PEDV infection and pathogenicity in suckling piglets.Palmitic acid(PA)was identified as one of those metabolites with metabolomic technology,and significantly enhanced PEDV replication in IPEC-J2 cells and PEDV infection and pathogenicity in suckling piglets.PA also increased the neutralizing antibody titer in the immune sera of mice.Furthermore,PA mediated the palmitoylation of the PEDV S protein,which improved virion stability and membrane fusion,thereby enhancing viral infection.Overall,our study demonstrates a novel mechanism of PEDV infection,with implications for PEDV pathogenicity.展开更多
Sepsis is a life-threatening condition caused by a dysregulated host response to infection,with its high mortality closely linked to excessive neutrophil activation and the release of neutrophil extracellular traps(NE...Sepsis is a life-threatening condition caused by a dysregulated host response to infection,with its high mortality closely linked to excessive neutrophil activation and the release of neutrophil extracellular traps(NETs).While NETs play a critical role in antimicrobial immunity,their overproduction can lead to tissue damage and multiple organ dysfunction,exacerbating the pathological progression of sepsis.This study focuses on the role of palmitoyltransferase ZDHHC18 in sepsis,proposing that it may regulate the formation of neutrophil NETs through palmitoylation,thereby promoting the onset of sepsis.Currently,there is no research,domestically or internationally,on the role of ZDHHC18 in NETs during sepsis.Therefore,exploring potential therapeutic strategies—such as inhibiting ZDHHC18 to mitigate the pathological effects of sepsis—could provide new insights for treating sepsis-related inflammatory diseases.展开更多
BACKGROUND Oral squamous cell carcinoma(OSCC)poses a major health burden,with frequent late-stage diagnoses and limited prognostic tools.Palmitoyl-protein thioesterase(PPT)1,a lysosomal enzyme,has been implicated in t...BACKGROUND Oral squamous cell carcinoma(OSCC)poses a major health burden,with frequent late-stage diagnoses and limited prognostic tools.Palmitoyl-protein thioesterase(PPT)1,a lysosomal enzyme,has been implicated in tumor biology.This study investigates PPT1 expression in OSCC and its association with clinicopathological features.AIM To evaluate the immunohistochemical expression of PPT1 in OSCC and assess its correlation with patient age,gender,tumor grade,and TNM staging using Hscore quantification.METHODS Immunohistochemistry for PPT1 was performed on 43 histopathologically confirmed OSCC samples.PPT1 expression was quantified using the H-score method(range:0-300).Correlation with clinical parameters was assessed using Spearman’s rank correlation and Kruskal-Wallis test.RESULTS PPT1 expression showed a significant positive correlation with tumor grade(ρ=0.48,P=0.0015),while a weak,nonsignificant negative correlation was noted with patient age(ρ=-0.27,P=0.083).No significant differences were found by gender or tumor stage.Given the small sample size,these findings should be interpreted as preliminary.CONCLUSION PPT1 expression is associated with histological tumor grade in OSCC and may reflect tumor aggressiveness.These initial results highlight the potential of PPT1 as a prognostic biomarker,warranting further validation in larger,multicenter studies.展开更多
基金supported by the National Natural Science Foundation of China (32473194)Natural Science Foundation of Zhejiang Province (LY23C190002)+1 种基金Natural Science Foundation of Ningbo City (202003N4011)One Health Interdisciplinary Research Project of Ningbo University (HZ202201)。
文摘NLRP3 inflammasome activation is pivotal for cytokine secretion and pyroptosis in response to diverse stimuli,playing a crucial role in innate immunity.While extensively studied in mammals,the regulatory mechanisms governing NLRP3 activation in non-mammalian vertebrates remain largely unexplored.Teleosts,as basal vertebrates,represent an ideal model for exploring the evolutionary trajectory of inflammasome regulation.In this study,ABE assays,confocal microscopy,and biochemical analyses were applied to systematically characterize the mechanisms underlying NLRP3 inflammasome in teleosts,using large yellow croakers(Larimichthys crocea,Lc)and zebrafish(Danio rerio,Dr)as representative models.Our findings revealed a previously unrecognized palmitoylation-dependent regulatory mechanism essential for teleost NLRP3 activation.Specifically,zDHHC18-mediated palmitoylation at a teleost-specific cysteine residue(C946 in LcNLRP3,C1037 in DrNLRP3)was required for the translocation of NLRP3 to the dispersed trans-Golgi network,facilitating its subsequent recruitment to the microtubule-organizing center.This membrane trafficking was crucial for inflammasome assembly and downstream inflammatory responses.These findings provide new insights into the distinct regulatory mechanisms of NLRP3 activation in teleosts,highlighting an evolutionary divergence that contributes to innate immunity adaptation in early vertebrates.
基金supported by Zhejiang Provincial Natural Science Foundation of China(Grant No.LZ25A040005)the National Natural Science Foundation of China(Grant No.11674287).
文摘The partitioning of membrane proteins into lipid domains in cellular membranes is closely associated with the realization of the protein functions and it is influenced by various factors such as the post-translational modification of palmitoylation.However,the molecular mechanism of the effect of palmitoylation on membrane protein partitioning into the lipid domains remains elusive.In this work,taking human peripheral myelin protein 22(PMP22)as an example,we employ coarse-grained molecular dynamics simulations to investigate the partitioning of both the natural PMP22 and the palmitoylated PMP22(pal-PMP22)into the lipid domains of model myelin membranes.The results indicate that palmitoylation drives PMP22 to localize at the boundary of the liquid-ordered(Lo)and liquid-disordered(Ld)domains and increases the possibility of PMP22 partitioning into the Lo domains by changing the hydrophobic length of the proteins and perturbing the ordered packing of tails of the saturated lipids in the Lo domains.This work offers some novel insights into the role of palmitoylation in modulating the function of membrane proteins in cellular membranes.
文摘Multicellular organisms use chemical messengers to transmit signals among organelles and to other cells. Relatively small hydrophobic molecules such as lipids are excellent candidates for this signaling purpose. In most proteins, palmitic acid and other saturated and some unsaturated fatty acids are esterified to the free thiol of cysteines and to the N-amide terminal. This palmitoylation process enhances the surface hydrophobicity and membrane affinity of protein substrates and plays important roles in modulating proteins’ trafficking, stability, and sorting etc. Protein palmitoylation has been involved in numerous cellular processes, including signaling, apoptosis, and neuronal transmission. The palmitoylation process is involved in multiple diseases such as Huntington’s disease, various cardiovascular and T-cell mediated immune disorders, as well as cancer. Protein palmitoylation through the thioester (S-acylation) is unique in that it is the only reversible lipid modification. Our study on lipopolysaccharide (LPS) and deoxynivalenol (DON) treatment to rats provides some insights to the complex role of protein palmitoylation in chemical and microbial toxicity. In contrast, myrisoylated proteins contain the 14-carbon fatty acid myristate attached via amide linkage to the N-terminal glycine residue of protein, and occur cotranslationally. The bacterial outer membrane enzyme lipid A palmitoyltransferase (PagP) confers resistance to host immune defenses by transferring a palmitate chain from a phospholipid to the lipid A component of LPS. PagP is sensitive to cationic antimicrobial peptides (CAMP) which are included among the products of the Toll-like receptor 4 (TLR4) signal transduction pathway. This modification of lipid A with a palmitate appears to both and protects the pathogenic bacteria from host immune defenses and attenuates the activation of those same defenses through the TLR4 signal transduction pathway.
基金supported by the Clinical Research Project of Investigator-Initiated Trials(IIT)Medical-Industry Integration Program of the Pudong New Area Health Commission(2025-PWYC-04)+1 种基金the Shanghai Oriental Talent Program(QNJY2024191)the National Natural Science Foundation of China(Youth Program,No.82303349).
文摘Palmitoylation plays a key role in liver cancer(HCC)initiation,progression,and drug resistance.This review highlights palmitoyltransferases(PATs),like ZDHHC family enzymes,and their impact on HCC and immune evasion.We discuss small-molecule inhibitors targeting these enzymes,such as ZDHHC9 and PPT1 inhibitors,showing promising results.The role of palmitoylation in immune checkpoint molecules like TIM-3 is explored,along with its potential in combination therapy.Additionally,we examine palmitoylation-related biomarkers for non-invasive diagnosis,prognosis,and treatment monitoring in liver cancer.
基金supported by the Natural Science Foundation of Shandong Province(grant No.ZR201911110516,China)the Taishan Scholar Program of Shandong Province(No.tsqn202408156,China).
文摘Palmitoylation,an essential covalent attachment of a fatty acid(usually C16 palmitate)to cysteine residues within proteins,is crucial for regulating protein functionality and enzymatic activities.This lipid modification facilitates the anchoring of proteins to cellular membranes,dictating their subcellular distribution and influencing protein transport dynamics and intracellular positioning.Additionally,it plays a role in regulating protein degradation through the ubiquitin-proteasome system.Palmitoylation is implicated in the pathogenesis and progression of cardiovascular diseases by modulating substrates and prompting additional post-translational modifications,as well as by interacting with other molecular alterations.Moreover,an intervention strategy focusing on palmitoylation processes is anticipated to offer novel therapeutic avenues for cardiovascular pathologies and address extant challenges in clinical settings.This review consolidates current research on the role and importance of palmitoylation in cardiovascular diseases by exploring its regulatory functions,the catalyzing enzymes,and the involved substrates.It highlights recent discoveries connecting palmitoylation-targeted therapies to cardiovascular health and examines potential approaches and future challenges in cardiovascular treatment.
基金supported by the National Natural Science Foundation of China(32225047)Double first-class discipline promotion project(2023B10564001)+2 种基金Open Project Program of Sichuan Provincial Key Laboratory of Animal Disease-resistant Nutrition,Sichuan Agricultural University(SZ202301-02)Guangdong Basic and Applied Basic Research Foundation(2022A1515111229)Hainan Provincial Natural Science Foundation of China(325QN483).
文摘Fatty acid metabolism mediates macrophage function;however,the underlying mechanism by which fatty acid metabolism regulates macrophage interleukin(IL)-1βproduction remains to be uncovered.Here,we used genome-wide association studies(GWAS)to identify several porcine serum IL-1β-related genes,such as the fatty acid metabolizing enzyme acyl-CoA thioesterase 11(ACOT11).We then demonstrated that inflammatory macrophages have low expression of ACOT11,and ACOT11 overexpression inhibits IL-1βmaturation from inflammatory macrophages.Mechanistically,ACOT11 promotes intracellular fatty acids accumulation,including eicosatetraenoic acid(EA)and stearic acid(SA),which inhibit activation of the Janus kinase(JAK)—signal transducer and activator of transcription(STAT)signaling through palmitoylation of interferon(IFN)-γreceptor(IFNGR)2 at C261site.Furthermore,we also found that EA attenuates lipopolysaccharide(LPS)-induced sepsis in mice.Collectively,our findings reveal a mechanism involving ACOT11-mediated post-translational modification that regulates macrophage function and provide a promising therapeutic target for the treatment of inflammatory diseases associated with macrophages.
基金supported in part by the National Key Research and Development Program of China(2023YFC3402100 to J.G.)the National Nature Science Foundation of China(82473404 to W.X.)the Natural Science Foundation of Guangdong Province(2022A1515220004 to J.G.,2024A1515010945 to W.X.,and 2023A1515010384 to Q.J.).
文摘Nutrients from dietary foods not only provide energy and building blocks,but also play critical roles in modulating diverse pathophysiological functions.They achieve these,in part,by accelerating cell signaling transduction processes via modulating various types of protein post-translational modifications(PTMs).Notably,accumulating evidence has identified palmitic acid(PA),a major component of high-fat diets,as a significant contributor to various human disorders,including diabetes and cancer.Hence,further understanding the roles of PA and its involvement in protein palmitoylation,a key PTM,is crucial for uncovering the mechanisms underlying these diseases and exploring potential clinical applications in cancer therapy.This review comprehensively summarizes recent advances in the understanding of PA homeostasis and palmitoylation in tumorigenesis.Specifically,it highlights the connections between palmitoylation and key processes such as oncogenic signaling pathways,cell death mechanisms,innate immune responses,and the tumor microenvironment.The review also emphasizes potential therapeutic strategies,including targeting PA homeostasis,palmitoylation-associated processes,or specific palmitoylated proteins for cancer treatment.Finally,the challenges in the field,such as the regulation of PA homeostasis and the dynamic detection or targeting of palmitoylation,are discussed,underscoring the need for further research to address these critical issues.
基金financially supported by the NIH R01 grant GM040602 (CAF)
文摘Protein palmitoylation is a widespread lipid modification in which one or more cysteine thiols on a substrate protein are modified to form a thioester with a palmitoyl group.This lipid modification is readily reversible;a feature of protein palmitoylation that allows for rapid regulation of the function of many cellular proteins.Mutations in palmitoyltransferases(PATs),the enzymes that catalyze the formation of this modification,are associated with a number of neurological diseases and cancer progression.This review summarizes the crucial role of palmitoylation in biological systems,the discovery of the DHHC protein family that catalyzes protein palmitoylation,and the development of methods for investigating the catalytic mechanism of PATs.
基金supported by the National Key Research and Development Program of China (2020YFA0803402 and2019YFA0801703)the National Natural Science Foundation of China(81872240,81802745,81790250/81790253 and 91959202)Innovation Program of Shanghai Municipal Education Commission (N173606)。
文摘Epithelial ovarian cancer(EOC) exhibits strong dependency on the tricarboxylic acid(TCA) cycle and oxidative phosphorylation to fuel anabolic process.Here,we show that malate dehydrogenase 2(MDH2),a key enzyme of the TCA cycle,is palmitoylated at cysteine 138(C138) residue,resulting in increased activity of MDH2.We next identify that ZDHHC18 acts as a palmitoyltransferase of MDH2.Glutamine deprivation enhances MDH2 palmitoylation by increasing the binding between ZDHHC18 and MDH2.MDH2 silencing represses mitochondrial respiration as well as ovarian cancer cell proliferation both in vitro and in vivo.Intriguingly,re-expression of wild-type MDH2,but not its palmitoylation-deficient C138 S mutant,sustains mitochondrial respiration and restores the growth as well as clonogenic capability of ovarian cancer cells.Notably,MDH2 palmitoylation level is elevated in clinical cancer samples from patients with high-grade serous ovarian cancer.These observations suggest that MDH2 palmitoylation catalyzed by ZDHHC18 sustains mitochondrial respiration and promotes the malignancy of ovarian cancer,yielding possibilities of targeting ZDHHC18-mediated MDH2 palmitoylation in the treatment of EOC.
基金supported by grants from the National Basic Research Program of China (the 973 Program, No. 2013CB531303 to Dr. Jianguo Chen No. 2014CB744601 to Fang Wang)+3 种基金the National Natural Scientific Foundation of China (NSFC, No. 81222048 to Fang Wang No. 81302754 to Peng Fei Wu)supported by the International Science & Technology Cooperation Program of China (No. 2011DFA32670 to Jianguo Chen)PCSIRT (No. IRT13016)
文摘Glutamate acting on AMPA-type ionotropic glutamate receptor(AMPAR) mediates the majority of fast excitatory synaptic transmission in the mammalian central nervous system. Dynamic regulation of AMPAR by post-translational modifications is one of the key elements that allow the nervous system to adapt to environment stimulations. S-palmitoylation, an important lipid modification by post-translational addition of a long-chain fatty acid to a cysteine residue, regulates AMPA receptor trafficking, which dynamically affects multiple fundamental brain functions, such as learning and memory. In vivo, S-palmitoylation is controlled by palmitoyl acyl transferases and palmitoyl thioesterases.In this review, we highlight advances in the mechanisms for dynamic AMPA receptors palmitoylation,and discuss how palmitoylation affects AMPA receptors function at synapses in recent years.Pharmacological regulation of S-palmitoylation may serve as a novel therapeutic strategy for neurobiological diseases.
文摘Gain-of-function mutations in the STING-encoding gene TMEM173 are central to the pathology of the autoinflammatory disorder STING-associated vasculopathy with onset in infancy(SAVI).Furthermore,excessive activity of the STING signaling pathway is associated with autoinflammatory diseases,including systemic lupus erythematosus and Aicardi–Goutières syndrome(AGS).Two independent studies recently identified pharmacological inhibitors of STING.Strikingly,both types of compounds are reactive nitrocontaining electrophiles that target STING palmitoylation,a posttranslational modification necessary for STING signaling.As a consequence,the activation of downstream signaling molecules and the induction of type I interferons were inhibited.The compounds were effective at ameliorating inflammation in a mouse model of AGS and in blocking the production of type I interferons in primary fibroblasts from SAVI patients.This mini-review focuses on the roles of palmitoylation in STING activation and signaling and as a pharmaceutical target for drug development.
基金the financial supports from the National Natural Science Foundation of China(91853205,81625022,81821005 to Cheng Luo81973166,21702218,91753207 to Bing Zhou),Wong Education to Cheng Luo and Bing Zhou+2 种基金the Department of Science and Technology of Fujian Province(2019T3029 to Cheng Luo,China)the Science and Technology Commission of Shanghai Municipality(19XD1404700 and 18431907100 to Cheng Luo,China)National Science&Technology Major Project“Key New Drug Creation and Manufacturing Program”(2018ZX09711002-008 to Cheng Luo,2018ZX09711002-008-005 to Huijin Feng,2018ZX09711002-006 to Bing Zhou,China)。
文摘The TEA domain(TEAD)family proteins(TEAD1-4)are essential transcription factors that control cell differentiation and organ size in the Hippo pathway.Although the sequences and structures of TEAD family proteins are highly conserved,each TEAD isoform has unique physiological and pathological functions.Therefore,the development and discovery of subtype selective inhibitors for TEAD protein will provide important chemical probes for the TEAD-related function studies in development and diseases.Here,we identified a novel TEAD 1/3 covalent inhibitor(DC-TEADin1072)with biochemical IC50 values of 0.61±0.02 and 0.58±0.12μmol/L against TEAD1 and TEAD3,respectively.Further chemical optimization based on DC-TEAD in 1072 yielded a selective TEAD3 inhibitor DCTEAD3 in03 with the IC_(50) value of 0.16±0.03μmol/L,which shows 100-fold selectivity over other TEAD isoforms in activity-based protein profiling(ABPP)assays.In cells,DC-TEAD3 in03 showed selective inhibitory effect on TEAD3 in GAL4-TEAD(1-4)reporter assays with the IC50 value of1.15μmol/L.When administered to zebrafish juveniles,experiments showed that DC-TEAD3 in03 reduced the growth rate of zebrafish caudal fins,indicating the importance of TEAD3 activity in controlling proportional growth of vertebrate appendages.
文摘The native Goα was purified from bovine brain cortex and palmitoylated in vitro. The in vitro palmitoylation site was the same as that in vivo. The internal palmitoylation of purified native Goα was found to be largely maintained. The apparent palmitoylation ratio was significantly increased after the Goα was treated with DTT. The GTPγS binding characteristic of Goα was not influenced by palmitoylation, however, the affinity for LUVs was increased dramatically. The in vitro palmitoylation model of Goα provides a better basis for studying the functional role of G protein palmitoylation in signal transduction.
基金supported by the key international cooperation projects of the National Natural Science Foundation of China(No.81820108004)the major projects of the Zhejiang Provincial Department of Science and Technology(No.2021C03123)the Pediatric Leukemia Diagnosis and Therapeutic Technology Research Center of Zhejiang Province(No.JBZX-201904).
文摘Acyl-CoA synthetase long chain family member 5(ACSL5),is a member of the acyl-CoA synthetases(ACSs)family that activates long chain fatty acids by catalyzing the synthesis of fatty acyl-CoAs.The dysregulation of ACSL5 has been reported in some cancers,such as glioma and colon cancers.However,little is known about the role of ACSL5 in acute myeloid leukemia(AML).We found that the expression of ACSL5 was higher in bone marrow cells from AML patients compared with that from healthy donors.ACSL5 level could serve as an independent prognostic predictor of the overall survival of AML patients.In AML cells,the ACSL5 knockdown inhibited cell growth both in vitro and in vivo.Mechanistically,the knockdown of ACSL5 suppressed the activation of the Wnt/β-catenin pathway by suppressing the palmitoylation modification of Wnt3a.Additionally,triacsin c,a pan-ACS family inhibitor,inhibited cell growth and robustly induced cell apoptosis when combined with ABT-199,the FDA approved BCL-2 inhibitor for AML therapy.Our results indicate that ACSL5 is a potential prognosis marker for AML and a promising pharmacological target for the treatment of molecularly stratified AML.
基金This work was supported by the Key Project of National Natural Science Foundation of China(No.81530006 to Ruibao Ren)Shanghai Collaborative Innovation Program on Regenerative Medicine and Stem Cell Research(No.2019CXJQ01 to Ruibao Ren)+2 种基金National Natural ScienceFoundation of China(No.81870112 to Ruibao Ren,No.81770171 to Bo Jiao,and No.81970134 to Ping Liu)Samuel Waxman Cancer Research Foundation(to Ruibao Ren)the Innovative Research Team of High-level Local Universities in Shanghai.
文摘More than 85%of patients with uveal melanoma(UM)carry a GNAQ or GNA11 mutation at a hotspot codon(Q209)that encodes G proteinαsubunit q/11 polypeptides(Gα_(q/11)).GNAQ/11 relies on palmitoylation for membrane association and signal transduction.Despite the palmitoylation of GNAQ/11 was discovered long before,its implication in UM remains unclear.Here,results of palmitoylation-targeted mutagenesis and chemical interference approaches revealed that the loss of GNAQ/11 palmitoylation substantially affected tumor cell proliferation and survival in UM cells.Palmitoylation inhibition through the mutation of palmitoylation sites suppressed GNAQ/11^(Q209L)-induced malignant transformation in NIH3T3 cells.Importantly,the palmitoylation-deficient oncogenic GNAQ/11 failed to rescue the cell death initiated by the knock down of endogenous GNAQ/11 oncogenes in UM cells,which are much more dependent on Gα_(q/11) signaling for cell survival and proliferation than other melanoma cells without GNAQ/11 mutations.Furthermore,the palmitoylation inhibitor,2-bromopalmitate,also specifically disrupted Gα_(q/11) downstream signaling by interfering with the MAPK pathway and BCL2 survival pathway in GNAQ/11-mutant UM cells and showed a notable synergistic effect when applied in combination with the BCL2 inhibitor,ABT-199,in vitro.The findings validate that GNAQ/11 palmitoylation plays a critical role in UM and may serve as a promising therapeutic target for GNAQ/11-driven UM.
基金supported by the Swedish Research Council(2017-01286,2020-01840)the Swedish state under the agreement between the Swedish government and the county councils(ALF-agreement)(ALFGBG721581)+2 种基金the Gustaf V 80-years fund(FAI-2018-0466)the IngaBritt and Arne Lundberg Foundation(LU2017-0076)the Novo Nordisk Foundation(26844).
文摘Membrane-initiated estrogen receptorα(mERα)signaling has been shown to affect bone mass in murine models.However,it remains unknown which cell types mediate the mERα-dependent effects on bone.In this study,we generated a novel mouse model with a conditional C451A mutation in Esr1,which enables selective knockout of the palmitoylation site essential for the membrane localization of ERα(C451A^(f/f)).First,we used Runx2-Cre mice to generate Runx2-C451A^(f/f)mice with conditional inactivation of mERαsignaling in Runx2-expressing osteoblast lineage cells.No significant changes were observed in body weight,weights of estrogen-responsive organs,or serum concentrations of estradiol between female Runx2-C451A^(f/f)and homozygous C451A^(f/f)littermate controls.High-resolution microcomputed tomography analysis showed a consistent decrease in cortical bone mass in the tibia,femur,and vertebra L5 of Runx2-C451A^(f/f)mice and three-point bending analysis of humerus revealed an impaired mechanical bone strength in Runx2-C451A^(f/f)female mice compared to controls.Additionally,primary osteoblast cultures from mice lacking mERαsignaling showed impaired differentiation compared to controls.
基金supported by the National Natural Science Foundation of China(U23A20236)the Key Research and Development Program,Guidance Projects of Heilongjiang Province,China(GZ20220029)。
文摘The host intestinal microbiota has emerged as the third element in the interactions between hosts and enteric viruses,and potentially affects the infection processes of enteric viruses.However,the interaction of porcine enteric coronavirus with intestinal microorganisms during infection remains unclear.In this study,we used 16S-rRNA-based Illumina NovaSeq high-throughput sequencing to identify the changes in the intestinal microbiota of piglets mediated by porcine epidemic diarrhea virus(PEDV)infection and the effects of the alterations in intestinal bacteria on PEDV infection and its molecular mechanisms.The intestinal microbiota of PEDV-infected piglets had significantly less diversity than the healthy group and different bacterial community characteristics.Among the altered intestinal bacteria,the relative abundance of Clostridium perfringens was significantly increased in the PEDV-infected group.A strain of C.perfringens type A,named DQ21,was successfully isolated from the intestines of healthy piglets.The metabolites of swine C.perfringens type A strain DQ21 significantly enhanced PEDV replication in porcine intestinal epithelial cell clone J2(IPEC-J2)cells,and PEDV infection and pathogenicity in suckling piglets.Palmitic acid(PA)was identified as one of those metabolites with metabolomic technology,and significantly enhanced PEDV replication in IPEC-J2 cells and PEDV infection and pathogenicity in suckling piglets.PA also increased the neutralizing antibody titer in the immune sera of mice.Furthermore,PA mediated the palmitoylation of the PEDV S protein,which improved virion stability and membrane fusion,thereby enhancing viral infection.Overall,our study demonstrates a novel mechanism of PEDV infection,with implications for PEDV pathogenicity.
文摘Sepsis is a life-threatening condition caused by a dysregulated host response to infection,with its high mortality closely linked to excessive neutrophil activation and the release of neutrophil extracellular traps(NETs).While NETs play a critical role in antimicrobial immunity,their overproduction can lead to tissue damage and multiple organ dysfunction,exacerbating the pathological progression of sepsis.This study focuses on the role of palmitoyltransferase ZDHHC18 in sepsis,proposing that it may regulate the formation of neutrophil NETs through palmitoylation,thereby promoting the onset of sepsis.Currently,there is no research,domestically or internationally,on the role of ZDHHC18 in NETs during sepsis.Therefore,exploring potential therapeutic strategies—such as inhibiting ZDHHC18 to mitigate the pathological effects of sepsis—could provide new insights for treating sepsis-related inflammatory diseases.
文摘BACKGROUND Oral squamous cell carcinoma(OSCC)poses a major health burden,with frequent late-stage diagnoses and limited prognostic tools.Palmitoyl-protein thioesterase(PPT)1,a lysosomal enzyme,has been implicated in tumor biology.This study investigates PPT1 expression in OSCC and its association with clinicopathological features.AIM To evaluate the immunohistochemical expression of PPT1 in OSCC and assess its correlation with patient age,gender,tumor grade,and TNM staging using Hscore quantification.METHODS Immunohistochemistry for PPT1 was performed on 43 histopathologically confirmed OSCC samples.PPT1 expression was quantified using the H-score method(range:0-300).Correlation with clinical parameters was assessed using Spearman’s rank correlation and Kruskal-Wallis test.RESULTS PPT1 expression showed a significant positive correlation with tumor grade(ρ=0.48,P=0.0015),while a weak,nonsignificant negative correlation was noted with patient age(ρ=-0.27,P=0.083).No significant differences were found by gender or tumor stage.Given the small sample size,these findings should be interpreted as preliminary.CONCLUSION PPT1 expression is associated with histological tumor grade in OSCC and may reflect tumor aggressiveness.These initial results highlight the potential of PPT1 as a prognostic biomarker,warranting further validation in larger,multicenter studies.
