Polypropylene(PP) accounts for approximately 28.0% of the global polyolefin market,valued at $243.4 billion in 2022.Known for its lightweight,chemical resistance,costeffectiveness,high strength and melting point,PP is...Polypropylene(PP) accounts for approximately 28.0% of the global polyolefin market,valued at $243.4 billion in 2022.Known for its lightweight,chemical resistance,costeffectiveness,high strength and melting point,PP is widely used in various applications.Its properties and applications are closely tied to its tacticity.One-dimensional(1D) conventional 13C NMR has been extensively utilized to analyze PP tacticity,but its low sensitivity and longer relaxation time remain drawbacks.Typically,analyzing a single PP sample requires around 9 h of NMR time.Using of a cryogenically cooled 10 mm NMR probe can significantly reduce this measurement time;however,its high cost makes it inaccessible for most NMR laboratories.While the refocused insensitive nuclei enhanced by polarization transfer(RINEPT) technique is well-known for enhancing NMR sensitivity,there are no published studies using 1D 13C RINEPT to quantify PP tacticity.Relaxation agents like chromium(Ⅲ) acetylacetonate(Cr(acac)_(3)) have also been used to reduce relaxation times in polyolefin NMR analyses.Here we introduce a straightforward and easily implementable 1D 13C NMR method for rapid PP tacticity quantification.This method combines Cr(acac)_(3),Bruker's existing RINEPT pulse sequence(ineptrd),and our recently published ~1H decoupling sequence(bi_waltz65_256 pl) to eliminate ^(1)H decoupling artifacts.It is worth noting that decoupling artifacts are always present.When the signal-to-noise ratio(SNR) is low,these artifacts are obscured by noise.For example,in some two-dimensional(2D) or three-dimensional(3D) NMR spectra,decoupling artifacts are barely visible because of the low SNR.However,when attempting to observe weak signals in ^(1)D spectra,increasing the sample concentration or the number of scans enhances the SNR,revealing the decoupling artifacts.The decoupling artifacts appeared superimpose with some other weak signals,affecting the measurements of signal intensities.Therefore,improved ~1H-decoupling methods are crucial for such data acquisitio n.This synergy results in a 9.4-to 9.7-fold sensitivity enhancement,equating to an 88-to 94-fold reduction(9.4^(2)≈88,9.7^(2)≈94) in NMR acquisition time compared to conventional 1D ^(13)C NMR experiment with Cr(acac)_(3).The time savings are even more substantial compared to experiments without Cr(acac)_(3).The faster and quantitative approach is accessible to researchers with or without cryoprobes.Beyond PP,this method can be applied to tacticity measurements of other polyolefins,such as polybutene,polyhexene and polyoctene.展开更多
Methane decomposition using nickel, copper, and aluminum （Ni：Cu/Al） and nickel, copper, potassium, and aluminum （Ni：Cu：K/Al） modified nano catalysts has been investigated for carbon fibers, hydrogen and hydroca...Methane decomposition using nickel, copper, and aluminum （Ni：Cu/Al） and nickel, copper, potassium, and aluminum （Ni：Cu：K/Al） modified nano catalysts has been investigated for carbon fibers, hydrogen and hydrocarbon production. X-ray photoelectron spectroscopy （XPS）, static secondary ion mass spectrometry （SSIMS）, thermal gravimetric analysis （TGA）, Fourier transform infrared （FT-IR）, secondary electron microscopy/X-ray energy dispersive （SEM-EDX）, and temperature programmed desorption （TPD） were used to depict the chemistry of the catalytic results. These techniques revealed the changes in surface morphology and structure of Ni, Cu, Al, and K, and formation of bimetallic and trimetallic surface cationic sites with different cationic species, which resulted in the production of graphitic form of pure carbon on Ni：Cu/Al catalyst. The addition of K has a marked effect on the product selectivity and reactivity of the catalyst system. K addition restricts the formation of carbon on the surface and increases the production of hydrogen and C2, C3 hydrocarbons during the catalytic reaction whereas no hydrocarbons are produced on the sample without K. This study completely maps the modified surface structure and its relationship with the catalytic behavior of both systems. The process provides a flexible route for the production of carbon fibers and hydrogen on Ni：Cu/Al catalyst and hydrogen along with hydrocarbons on Ni：Cu：K/Al catalyst. The produced carbon fibers are imaged using a transmission electron microscope （TEM） for diameter size and wall structure determination. Hydrogen produced is COx free, which can be used directly in the fuel cell system. The effect of the addition of Cu and its transformation and interaction with Ni and K is responsible for the production of CO/CO2 free hydrogen, thus producing an environmental friendly clean energy.展开更多
Drug discovery is a crucial part of human healthcare and has dramatically benefited human lifespan and life quality in recent centuries, however, it is usually time-and effort-consuming. Structural biology has been de...Drug discovery is a crucial part of human healthcare and has dramatically benefited human lifespan and life quality in recent centuries, however, it is usually time-and effort-consuming. Structural biology has been demonstrated as a powerful tool to accelerate drug development. Among different techniques, cryo-electron microscopy(cryo-EM) is emerging as the mainstream of structure determination of biomacromolecules in the past decade and has received increasing attention from the pharmaceutical industry. Although cryo-EM still has limitations in resolution, speed and throughput, a growing number of innovative drugs are being developed with the help of cryo-EM. Here, we aim to provide an overview of how cryo-EM techniques are applied to facilitate drug discovery. The development and typical workflow of cryo-EM technique will be briefly introduced, followed by its specific applications in structure-based drug design, fragment-based drug discovery, proteolysis targeting chimeras, antibody drug development and drug repurposing. Besides cryo-EM, drug discovery innovation usually involves other state-of-the-art techniques such as artificial intelligence(AI), which is increasingly active in diverse areas. The combination of cryo-EM and AI provides an opportunity to minimize limitations of cryo-EM such as automation, throughput and interpretation of mediumresolution maps, and tends to be the new direction of future development of cryo-EM. The rapid development of cryo-EM will make it as an indispensable part of modern drug discovery.展开更多
Octameric hemoglobins have been developed by the introduction of surface cysteines in either the alpha or beta chain. Originally designed as a blood substitute, we report here the structure and ligand binding function...Octameric hemoglobins have been developed by the introduction of surface cysteines in either the alpha or beta chain. Originally designed as a blood substitute, we report here the structure and ligand binding function;in addition the interaction with haptoglobin was studied. The recombinant Hbs (rHbs) with mutations alpha Asn78Cys or beta Gly83Cys spontaneously form octamers under conditions where the cysteines are oxidized. Oxygen binding curves and CO kinetic studies indicate a correct allosteric transition of the tetramers within the octamer. Crystallographic studies of the two rHbs show two disulfide bonds per octamer. Reducing agents may provoke dissociation to tetramers, but the octamers are stable when mixed with fresh human plasma, indicating that the reduction by plasma is slower than the oxidation by the dissolved oxygen, consistent with an enhanced stability. The octameric rHbs were also mixed with a solution of haptoglobin (Hp), which binds the dimers of Hb: there was little interaction for incubation times of 15 min;however, on longer timescales a complex was formed. Dynamic light scattering was used to follow the interaction of Hp with the alpha Asn78Cys octamer during 24 hours;a transition from a simple complex of 15 nm to a final size of 60 nm was observed. The results indicate a specific orientation of the αβ dimers may be of importance for the binding to haptoglobin.展开更多
Fragile X syndrome(FXS)is the most prevalent inherited intellectual disability,resulting from a loss of fragile X mental retardation protein(FMRP).Patients with FXS suffer lifelong cognitive disabilities,but the funct...Fragile X syndrome(FXS)is the most prevalent inherited intellectual disability,resulting from a loss of fragile X mental retardation protein(FMRP).Patients with FXS suffer lifelong cognitive disabilities,but the function of FMRP in the adult brain and the mechanism underlying age-related cognitive decline in FXS is not fully understood.Here,we report that a loss of FMRP results in increased protein synthesis of histone acetyltransferase EP300 and ubiquitinationmediated degradation of histone deacetylase HDAC1 in adult hippocampal neural stem cells(NSCs).Consequently,FMRPdeficient NSCs exhibit elevated histone acetylation and age-related NSC depletion,leading to cognitive impairment in mature adult mice.Reducing histone acetylation rescues both neurogenesis and cognitive deficits in mature adult FMRPdeficient mice.Our work reveals a role for FMRP and histone acetylation in cognition and presents a potential novel ther⁃apeutic strategy for treating adult FXS patients.展开更多
AIM:To determine the effect of pituitary adenylate cy-clase-activating polypeptide (PACAP) on left gastric artery (LGA) flow and to unveil the structural or functional important sites that may be critical for discrimi...AIM:To determine the effect of pituitary adenylate cy-clase-activating polypeptide (PACAP) on left gastric artery (LGA) flow and to unveil the structural or functional important sites that may be critical for discrimination of different receptor subtypes. METHODS: Peptides, including PACAP-27, PACAP-38, amino acid substituted PACAP-27 and C-terminus truncated analogues PACAP (27-38), were synthesized by a simultaneous multiple solid-phase peptide synthesizer. Flow probes of an ultrasound transit-time blood flowmeter were placed around the LGA of beagle dogs. Whenpeptides were infused intravenously, the blood flow was measured.RESULTS: [Ala4, Val5]-PACAP-27 caused a concentration-dependent vasodepressor action which was similar to that caused by PACAP-27. The LGA blood flow response to [Ala4, Val5]-PACAP-27 was significantly higher than that to PACAP-27, which was similar to that to vasoactive intestinal polypeptide (VIP) at the same dose. [Ala6]-PACAP-27 did not increase the peak LGA ? ow. [Gly8]-PACAP-27 showed a similar activity to VIP. [Asn24, Ser25, Ile26]-PACAP-27 did not change the activity of peptides at all doses. CONCLUSION: NH2 terminus is more important to biological activity of peptides and specifi c receptor recognition than COOH-terminus.展开更多
AIM:To identify non-quinazoline kinase inhibitors effective against drug resistant mutants of epidermal growth factor receptor(EGFR).METHODS:A kinase inhibitor library was subjected to screening for specific inhibitio...AIM:To identify non-quinazoline kinase inhibitors effective against drug resistant mutants of epidermal growth factor receptor(EGFR).METHODS:A kinase inhibitor library was subjected to screening for specific inhibition pertaining to the in vitro kinase activation of EGFR with the gatekeeper mutation T790 M,which is resistant to small molecular weight tyrosine kinase inhibitors(TKIs) for EGFR in nonsmall cell lung cancers(NSCLCs). This inhibitory effect was confirmed by measuring autophosphorylation of EGFR T790M/L858 R in NCI-H1975 cells,an NSCLC cell line harboring the gatekeeper mutation. The effects of a candidate compound,Janus kinase 3(JAK3) inhibitor Ⅵ,on cell proliferation were evaluated using the MTT assay and were compared between T790M-positive and-negative lung cancer cell lines. JAK3 inhibitor Ⅵ was modeled into the ATP-binding pocket of EGFR T790M/L858 R. Potential physical interactions between the compound and kinase domains of wild-type(WT) or mutant EGFRs or JAK3 were estimated by calculating binding energy. The gatekeeper residues of EGFRs and JAKs were aligned to discuss the similarities among EGFR T790 M and JAKs. RESULTS:We found that JAK3 inhibitor Ⅵ,a known inhibitor for JAK3 tyrosine kinase,selectively inhibits EGFR T790M/L858 R,but has weaker inhibitory effects on the WT EGFR in vitro. JAK3 inhibitor Ⅵ also specifically reduced autophosphorylation of EGFR T790M/L858 R in NCI-H1975 cells upon EGF stimulation,but did not show the inhibitory effect on WT EGFR in A431 cells. Furthermore,JAK3 inhibitor Ⅵ suppressed the proliferationof NCI-H1975 cells,but showed limited inhibitory effects on the WT EGFR-expressing cell lines A431 and A549.A docking simulation between JAK3 inhibitor Ⅵ and the ATP-binding pocket of EGFR T790M/L858 R predicted a potential binding status with hydrogen bonds. Estimated binding energy of JAK3 inhibitor Ⅵ to EGFR T790M/L858 R was more stable than its binding energy to the WT EGFR. Amino acid sequence alignments revealed that the gatekeeper residues of JAK family kinases are methionine in WT,similar to EGFR T790 M,suggesting that TKIs for JAKs may also be effective for EGFR T790 M. CONCLUSION:Our findings demonstrate that JAK3 inhibitor Ⅵ is a gatekeeper mutant selective TKI and offer a strategy to search for new EGFR T790 M inhibitors.展开更多
Staphylococcus aureus is pathogenic to humans with worldwide health care concern due to its ability to evade the immune system and develop resistance to multiple drugs. Reg family proteins are C-type lectins with anti...Staphylococcus aureus is pathogenic to humans with worldwide health care concern due to its ability to evade the immune system and develop resistance to multiple drugs. Reg family proteins are C-type lectins with antimicrobial properties. Bacterial aggregation through binding to microbial cell surface sugar and/or lipid moieties is key mechanism employed in the process. In the present study we have analysed the antimicrobial effect of human REG Iα on S. aureus. Aggregation of mid-log phase culture of S. aureus was observed in presence of purified recombinant REG Iα. Therefore REG Iα can be applied in eliminating S. aureus infections in humans.展开更多
Riboflavin （Rf） receptors bind and translocate Rf and its phosphorylated forms （e.g. flavin mononucleotide, FMN） into cells where they mediate various cellular metabolic pathways. Previously, we showed that FMN-co...Riboflavin （Rf） receptors bind and translocate Rf and its phosphorylated forms （e.g. flavin mononucleotide, FMN） into cells where they mediate various cellular metabolic pathways. Previously, we showed that FMN-coated ultrasmall superparamagnetic iron oxide （FLUSPIO） nanoparticles are suitable for labeling metabolically active cancer and endothelial cells in vitro. In this study, we focused on the in vivo application of FLUSPIO using prostate cancer xenografts. Size, charge, and chemical composition of FLUSPIO were evaluated. We explored the in vitro specificity of FLUSPIO for its cellular receptors using magnetic resonance imaging （MRI） and Prussian blue staining. Competitive binding experiments were performed in vivo by injecting free FMN in excess. Bio-distribution of FLUSPIO was determined by estimating iron content in organs and tumors using a colorimetric assay. AFM analysis and zeta potential measurements revealed a particulate morphology approximately 20-40 nm in size and a negative zeta potential （-24.23±0.15 mV） in water. X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry data confirmed FMN present on the USPIO nanoparticle surface. FLUSPIO uptake in prostate cancer cells and human umbilical vein endothelial cells was significantly higher than that of control USPIO, while addition of excess of free FMN reduced accumulation. Similarly, in vivo MRI and histology showed specific FLUSPIO uptake by prostate cancer cells, tumor endothelial cells, and tumor-associated macrophages. Besides prominent tumor accumulation, FLUSPIO accumulated in the liver, spleen, lung, and skin. Hence, our data strengthen our hypothesis that targeting riboflavin receptors is an efficient approach to accumulate nanomedicines in tumors opening perspectives for the development of diagnostic and therapeutic systems.展开更多
Jasmonates(JAs)are plant hormones with crucial roles in development and stress resilience.They activate MYC transcription factors by mediating the proteolysis of MYC inhibitors called JAZ proteins.In the absence of JA...Jasmonates(JAs)are plant hormones with crucial roles in development and stress resilience.They activate MYC transcription factors by mediating the proteolysis of MYC inhibitors called JAZ proteins.In the absence of JA,JAZ proteins bind and inhibit MYC through the assembly of MYC–JAZ–Novel Interactor of JAZ(NINJA)–TPL repressor complexes.However,JAZ and NINJA are predicted to be largely intrinsically unstructured,which has precluded their experimental structure determination.Through a combination of biochemical,mutational,and biophysical analyses and AlphaFold-derived ColabFold modeling,we characterized JAZ–JAZ and JAZ–NINJA interactions and generated models with detailed,high-confidence domain interfaces.We demonstrate that JAZ,NINJA,and MYC interface domains are dynamic in isolation and become stabilized in a stepwise order upon complex assembly.By contrast,most JAZ and NINJA regions outside of the interfaces remain highly dynamic and cannot be modeled in a single conformation.Our data indicate that the small JAZ Zinc finger expressed in Inflorescence Meristem(ZIM)motif mediates JAZ–JAZ and JAZ–NINJA interactions through separate surfaces,and our data further suggest that NINJA modulates JAZ dimerization.This study advances our understanding of JA signaling by providing insights into the dynamics,interactions,and structure of the JAZ–NINJA core of the JA repressor complex.展开更多
Wound healing is a complex and multistep biological process that involves the cooperation of various cell types.Programmed cell death,including apoptosis and necrotizing apoptosis,plays a crucial role in this process....Wound healing is a complex and multistep biological process that involves the cooperation of various cell types.Programmed cell death,including apoptosis and necrotizing apoptosis,plays a crucial role in this process.Apoptosis,a controlled and orderly programmed cell death regulated by genes,helps eliminate unnecessary or abnormal cells and maintain internal environmental stability.It also regulates various cell functions and contributes to the development of many diseases.In wound healing,programmed cell death is essential for removing inflammatory cells and forming scars.On the other hand,necroptosis,another form of programmed cell death,has not been thoroughly investigated regarding its role in wound healing.This review explores the changes and apoptosis of specific cell groups during wound healing after an injury and delves into the potential underlying mechanisms.Furthermore,it briefly discusses the possible mechanisms linking wound inflammation and fibrosis to apoptosis in wound healing.By understanding the relationship between apoptosis and wound healing and investigating the molecular mechanisms involved in apoptosis regulation,new strategies for the clinical treatment of wound healing may be discovered.展开更多
Bacterial infection of implanted scaffolds may have fatal consequences and,in combination with the emergence of multidrug bacterial resistance,the development of advanced antibacterial biomaterials and constructs is o...Bacterial infection of implanted scaffolds may have fatal consequences and,in combination with the emergence of multidrug bacterial resistance,the development of advanced antibacterial biomaterials and constructs is of great interest.Since decades ago,metals and their ions had been used to minimize bacterial infection risk and,more recently,metal-based nanomaterials,with improved antimicrobial properties,have been advocated as a novel and tunable alternative.A comprehensive review is provided on how metal ions and ion nanoparticles have the potential to decrease or eliminate unwanted bacteria.Antibacterial mechanisms such as oxidative stress induction,ion release and disruption of biomolecules are currently well accepted.However,the exact antimicrobial mechanisms of the discussed metal compounds remain poorly understood.The combination of different metal ions and surface decorations of nanoparticles will lead to synergistic effects and improved microbial killing,and allow to mitigate potential side effects to the host.Starting with a general overview of antibacterial mechanisms,we subsequently focus on specific metal ions such as silver,zinc,copper,iron and gold,and outline their distinct modes of action.Finally,we discuss the use of these metal ions and nanoparticles in tissue engineering to prevent implant failure.展开更多
CYP3A5 is a cytochrome P450(CYP)enzyme that metabolizes drugs and contributes to drug resistance in cancer.However,it remains unclear whether CYP3A5 directly influences cancer progression.In this report,we demonstrate...CYP3A5 is a cytochrome P450(CYP)enzyme that metabolizes drugs and contributes to drug resistance in cancer.However,it remains unclear whether CYP3A5 directly influences cancer progression.In this report,we demonstrate that CYP3A5 regulates glucose metabolism in pancreatic ductal adenocarcinoma.Multi-omics analysis showed that CYP3A5 knockdown re-sults in a decrease in various glucose-related metabolites through its effect on glucose trans-port.A mechanistic study revealed that CYP3A5 enriches the glucose transporter GLUT1 at the plasma membrane by restricting the translation of TXNIP,a negative regulator of GLUT1.Notably,CYP3A5-generated reactive oxygen species were proved to be responsible for atten-uating the AKT-4EBP1-TXNIP signaling pathway.CYP3A5 contributes to cell migration by maintaining high glucose uptake in pancreatic cancer.Taken together,our results,for the first time,reveal a role of CYP3A5 in glucose metabolism in pancreatic ductal adenocarcinoma and identify a novel mechanism that is a potential therapeutic target.展开更多
Precise and high-speed sorting of individual target cells from heterogeneous populations plays an imperative role in cell research.Although the conventional fluorescence-activated cell sorter(FACs)is capable of rapid ...Precise and high-speed sorting of individual target cells from heterogeneous populations plays an imperative role in cell research.Although the conventional fluorescence-activated cell sorter(FACs)is capable of rapid and accurate cell sorting,it occupies a large volume of the instrument and inherently brings in aerosol generation as well as crosscontamination among samples.The sorting completed in a fully enclosed and disposable microfluidic chip has the potential to eliminate the above concerns.However,current microfluidic cell sorters are hindered by the high.complexities of the fabrication procedure and the off-chip setup.In this paper,a spark-cavitation-bubble-based fluorescence-activated cell sorter is developed to perform fast and accurate sorting in a microfluidic chip.It features a simple structure and an easy operation.This microfluidic sorter comprises a positive electrode of platinum and a negative electrode of tungsten,which are placed on the side of the main channel.By applying a high-voltage discharge on the pair of electrodes,a single spark cavitation bubble is created to deflect the target particle into the downstream collection channel.The sorter has a short switching time of 150μs and a long lifespan of more than 100 million workable actions.In addition,a novel control strategy is proposed to dynamically adjust the discharge time to stabilize the size of the cavitation bubble for continuous sorting.The dynamic control of continuously triggering the sorter,the optimal delay time between fluorescence detection and cell sorting,and a theoretical model to predict the ideal sorting recovery and purity are studied to improve and evaluate the sorter performance.The experiments demonstrate that the sorting rate of target particles achieves 1200 eps,the total analysis throughput is up to 10,000 eps,the particles sorted at 4000 eps exhibit a purity greater than 80%and a recovery rate greater than 90%,and the sorting effect on the viability of HelLa cells is negligible.展开更多
Optical imaging techniques provide low-cost,non-radiative images with high spatiotemporal resolution,making them advantageous for long-term dynamic observation of blood perfusion in stroke research and other brain stu...Optical imaging techniques provide low-cost,non-radiative images with high spatiotemporal resolution,making them advantageous for long-term dynamic observation of blood perfusion in stroke research and other brain studies compared to non-optical methods.However,high-resolution imaging in optical microscopy fundamentally requires a tight optical focus,and thus a limited depth of field(DOF).Consequently,large-scale,non-stitched,high-resolution images of curved surfaces,like brains,are difficult to acquire without z-axis scanning.To overcome this limitation,we developed a needle-shaped beam optical coherence tomography angiography(NB-OCTA)system,and for the first time,achieved a volumetric resolution of less than 8μm in a non-stitched volume space of 6.4 mm×4 mm×620μm in vivo.This system captures the distribution of blood vessels at 3.4-times larger depths than normal OCTA equipped with a Gaussian beam(GB-OCTA).We then employed NB-OCTA to perform long-term observation of cortical blood perfusion after stroke in vivo,and quantitatively analyzed the vessel area density(VAD)and the diameters of representative vessels in different regions over 10 days,revealing different spatiotemporal dynamics in the acute,sub-acute and chronic phase of post-ischemic revascularization.Benefiting from our NB-OCTA,we revealed that the recovery process is not only the result of spontaneous reperfusion,but also the formation of new vessels.This study provides visual and mechanistic insights into strokes and helps to deepen our understanding of the spontaneous response of brain after stroke.展开更多
Proteins play a critical role in biology and biopharma due to their specificity and minimal side effects.Predicting the effects of mutations on protein stability is vital but experimentally challenging.Deep learning o...Proteins play a critical role in biology and biopharma due to their specificity and minimal side effects.Predicting the effects of mutations on protein stability is vital but experimentally challenging.Deep learning offers an efficient solution to this problem.In the present work,we introduced ProstaNet,a deep learning framework that predicts stability changes resulting from single-and multiple-point mutations using geometric vector perceptrons-graph neural network for 3-dimensional feature processing.For training ProstaNet,we meticulously crafted ProstaDB,a comprehensive and pristine thermodynamics repository,including 3,784 single-point mutations and 1,642 multiple-point mutations.We also created thermodynamic looping for enlarging the limited data size of multiple-point mutation and applied an innovative clustering method to generate a standard testing set of multiple-point mutation.Besides,we identified residue scoring as the most important encoding method in protein properties prediction.With these innovations,ProstaNet accurately predicts thermostability changes for both single-point and multiple-point mutations without showing any bias.ProstaNet achieves an accuracy of 0.75,outperforming existing methods for single-point mutation prediction,including ThermoMPNN(0.63),PoPMuSiC^(sym)(0.66),MUPRO(0.52),and FoldX(0.71).ProstaNet also achieves a 1.3-fold increase in accuracy compared to FoldX for multiple-point mutation predictions.Validated by experiment,4 out of 5 single-point mutation predictions(80%)and all multiple-point mutation predictions(100%)for HuJ3 mutants were accurate,demonstrating the potential benefits of ProstaNet for protein engineering and drug development.展开更多
文摘Polypropylene(PP) accounts for approximately 28.0% of the global polyolefin market,valued at $243.4 billion in 2022.Known for its lightweight,chemical resistance,costeffectiveness,high strength and melting point,PP is widely used in various applications.Its properties and applications are closely tied to its tacticity.One-dimensional(1D) conventional 13C NMR has been extensively utilized to analyze PP tacticity,but its low sensitivity and longer relaxation time remain drawbacks.Typically,analyzing a single PP sample requires around 9 h of NMR time.Using of a cryogenically cooled 10 mm NMR probe can significantly reduce this measurement time;however,its high cost makes it inaccessible for most NMR laboratories.While the refocused insensitive nuclei enhanced by polarization transfer(RINEPT) technique is well-known for enhancing NMR sensitivity,there are no published studies using 1D 13C RINEPT to quantify PP tacticity.Relaxation agents like chromium(Ⅲ) acetylacetonate(Cr(acac)_(3)) have also been used to reduce relaxation times in polyolefin NMR analyses.Here we introduce a straightforward and easily implementable 1D 13C NMR method for rapid PP tacticity quantification.This method combines Cr(acac)_(3),Bruker's existing RINEPT pulse sequence(ineptrd),and our recently published ~1H decoupling sequence(bi_waltz65_256 pl) to eliminate ^(1)H decoupling artifacts.It is worth noting that decoupling artifacts are always present.When the signal-to-noise ratio(SNR) is low,these artifacts are obscured by noise.For example,in some two-dimensional(2D) or three-dimensional(3D) NMR spectra,decoupling artifacts are barely visible because of the low SNR.However,when attempting to observe weak signals in ^(1)D spectra,increasing the sample concentration or the number of scans enhances the SNR,revealing the decoupling artifacts.The decoupling artifacts appeared superimpose with some other weak signals,affecting the measurements of signal intensities.Therefore,improved ~1H-decoupling methods are crucial for such data acquisitio n.This synergy results in a 9.4-to 9.7-fold sensitivity enhancement,equating to an 88-to 94-fold reduction(9.4^(2)≈88,9.7^(2)≈94) in NMR acquisition time compared to conventional 1D ^(13)C NMR experiment with Cr(acac)_(3).The time savings are even more substantial compared to experiments without Cr(acac)_(3).The faster and quantitative approach is accessible to researchers with or without cryoprobes.Beyond PP,this method can be applied to tacticity measurements of other polyolefins,such as polybutene,polyhexene and polyoctene.
文摘Methane decomposition using nickel, copper, and aluminum （Ni：Cu/Al） and nickel, copper, potassium, and aluminum （Ni：Cu：K/Al） modified nano catalysts has been investigated for carbon fibers, hydrogen and hydrocarbon production. X-ray photoelectron spectroscopy （XPS）, static secondary ion mass spectrometry （SSIMS）, thermal gravimetric analysis （TGA）, Fourier transform infrared （FT-IR）, secondary electron microscopy/X-ray energy dispersive （SEM-EDX）, and temperature programmed desorption （TPD） were used to depict the chemistry of the catalytic results. These techniques revealed the changes in surface morphology and structure of Ni, Cu, Al, and K, and formation of bimetallic and trimetallic surface cationic sites with different cationic species, which resulted in the production of graphitic form of pure carbon on Ni：Cu/Al catalyst. The addition of K has a marked effect on the product selectivity and reactivity of the catalyst system. K addition restricts the formation of carbon on the surface and increases the production of hydrogen and C2, C3 hydrocarbons during the catalytic reaction whereas no hydrocarbons are produced on the sample without K. This study completely maps the modified surface structure and its relationship with the catalytic behavior of both systems. The process provides a flexible route for the production of carbon fibers and hydrogen on Ni：Cu/Al catalyst and hydrogen along with hydrocarbons on Ni：Cu：K/Al catalyst. The produced carbon fibers are imaged using a transmission electron microscope （TEM） for diameter size and wall structure determination. Hydrogen produced is COx free, which can be used directly in the fuel cell system. The effect of the addition of Cu and its transformation and interaction with Ni and K is responsible for the production of CO/CO2 free hydrogen, thus producing an environmental friendly clean energy.
基金funded by the National Natural Science Foundation of China (NSFC, 31900046, 81972085, 82172465 and 32161133022)the Guangdong Provincial Key Laboratory of Advanced Biomaterials (2022B1212010003)+7 种基金the National Science and Technology Innovation 2030 Major Program (2022ZD0211900)the Shenzhen Key Laboratory of Computer Aided Drug Discovery (ZDSYS20201230165400001)the Chinese Academy of Science President’s International Fellowship Initiative (PIFI)(2020FSB0003)the Guangdong Retired Expert (granted by Guangdong Province)the Shenzhen Pengcheng ScientistNSFC-SNSF Funding (32161133022)Alpha Mol&SIAT Joint LaboratoryShenzhen Government Top-talent Working Funding and Guangdong Province Academician Work Funding。
文摘Drug discovery is a crucial part of human healthcare and has dramatically benefited human lifespan and life quality in recent centuries, however, it is usually time-and effort-consuming. Structural biology has been demonstrated as a powerful tool to accelerate drug development. Among different techniques, cryo-electron microscopy(cryo-EM) is emerging as the mainstream of structure determination of biomacromolecules in the past decade and has received increasing attention from the pharmaceutical industry. Although cryo-EM still has limitations in resolution, speed and throughput, a growing number of innovative drugs are being developed with the help of cryo-EM. Here, we aim to provide an overview of how cryo-EM techniques are applied to facilitate drug discovery. The development and typical workflow of cryo-EM technique will be briefly introduced, followed by its specific applications in structure-based drug design, fragment-based drug discovery, proteolysis targeting chimeras, antibody drug development and drug repurposing. Besides cryo-EM, drug discovery innovation usually involves other state-of-the-art techniques such as artificial intelligence(AI), which is increasingly active in diverse areas. The combination of cryo-EM and AI provides an opportunity to minimize limitations of cryo-EM such as automation, throughput and interpretation of mediumresolution maps, and tends to be the new direction of future development of cryo-EM. The rapid development of cryo-EM will make it as an indispensable part of modern drug discovery.
文摘Octameric hemoglobins have been developed by the introduction of surface cysteines in either the alpha or beta chain. Originally designed as a blood substitute, we report here the structure and ligand binding function;in addition the interaction with haptoglobin was studied. The recombinant Hbs (rHbs) with mutations alpha Asn78Cys or beta Gly83Cys spontaneously form octamers under conditions where the cysteines are oxidized. Oxygen binding curves and CO kinetic studies indicate a correct allosteric transition of the tetramers within the octamer. Crystallographic studies of the two rHbs show two disulfide bonds per octamer. Reducing agents may provoke dissociation to tetramers, but the octamers are stable when mixed with fresh human plasma, indicating that the reduction by plasma is slower than the oxidation by the dissolved oxygen, consistent with an enhanced stability. The octameric rHbs were also mixed with a solution of haptoglobin (Hp), which binds the dimers of Hb: there was little interaction for incubation times of 15 min;however, on longer timescales a complex was formed. Dynamic light scattering was used to follow the interaction of Hp with the alpha Asn78Cys octamer during 24 hours;a transition from a simple complex of 15 nm to a final size of 60 nm was observed. The results indicate a specific orientation of the αβ dimers may be of importance for the binding to haptoglobin.
文摘Fragile X syndrome(FXS)is the most prevalent inherited intellectual disability,resulting from a loss of fragile X mental retardation protein(FMRP).Patients with FXS suffer lifelong cognitive disabilities,but the function of FMRP in the adult brain and the mechanism underlying age-related cognitive decline in FXS is not fully understood.Here,we report that a loss of FMRP results in increased protein synthesis of histone acetyltransferase EP300 and ubiquitinationmediated degradation of histone deacetylase HDAC1 in adult hippocampal neural stem cells(NSCs).Consequently,FMRPdeficient NSCs exhibit elevated histone acetylation and age-related NSC depletion,leading to cognitive impairment in mature adult mice.Reducing histone acetylation rescues both neurogenesis and cognitive deficits in mature adult FMRPdeficient mice.Our work reveals a role for FMRP and histone acetylation in cognition and presents a potential novel ther⁃apeutic strategy for treating adult FXS patients.
基金Supported by (in part) Grants from Ministry of Education,Culture,Science,and Technology,Japan Society for the Promotion of Science and Special Fund of Six-Talented Peak of Jiangsu Province,No.07-B-15 (IB07)
文摘AIM:To determine the effect of pituitary adenylate cy-clase-activating polypeptide (PACAP) on left gastric artery (LGA) flow and to unveil the structural or functional important sites that may be critical for discrimination of different receptor subtypes. METHODS: Peptides, including PACAP-27, PACAP-38, amino acid substituted PACAP-27 and C-terminus truncated analogues PACAP (27-38), were synthesized by a simultaneous multiple solid-phase peptide synthesizer. Flow probes of an ultrasound transit-time blood flowmeter were placed around the LGA of beagle dogs. Whenpeptides were infused intravenously, the blood flow was measured.RESULTS: [Ala4, Val5]-PACAP-27 caused a concentration-dependent vasodepressor action which was similar to that caused by PACAP-27. The LGA blood flow response to [Ala4, Val5]-PACAP-27 was significantly higher than that to PACAP-27, which was similar to that to vasoactive intestinal polypeptide (VIP) at the same dose. [Ala6]-PACAP-27 did not increase the peak LGA ? ow. [Gly8]-PACAP-27 showed a similar activity to VIP. [Asn24, Ser25, Ile26]-PACAP-27 did not change the activity of peptides at all doses. CONCLUSION: NH2 terminus is more important to biological activity of peptides and specifi c receptor recognition than COOH-terminus.
基金the Screening Committee of Anticancer Drugs in the Scientific Support Programs for Cancer Research Grant--in--Aid for Scientific Research on Innovative Areas from the Ministry of Education,Culture,Sports,Science
文摘AIM:To identify non-quinazoline kinase inhibitors effective against drug resistant mutants of epidermal growth factor receptor(EGFR).METHODS:A kinase inhibitor library was subjected to screening for specific inhibition pertaining to the in vitro kinase activation of EGFR with the gatekeeper mutation T790 M,which is resistant to small molecular weight tyrosine kinase inhibitors(TKIs) for EGFR in nonsmall cell lung cancers(NSCLCs). This inhibitory effect was confirmed by measuring autophosphorylation of EGFR T790M/L858 R in NCI-H1975 cells,an NSCLC cell line harboring the gatekeeper mutation. The effects of a candidate compound,Janus kinase 3(JAK3) inhibitor Ⅵ,on cell proliferation were evaluated using the MTT assay and were compared between T790M-positive and-negative lung cancer cell lines. JAK3 inhibitor Ⅵ was modeled into the ATP-binding pocket of EGFR T790M/L858 R. Potential physical interactions between the compound and kinase domains of wild-type(WT) or mutant EGFRs or JAK3 were estimated by calculating binding energy. The gatekeeper residues of EGFRs and JAKs were aligned to discuss the similarities among EGFR T790 M and JAKs. RESULTS:We found that JAK3 inhibitor Ⅵ,a known inhibitor for JAK3 tyrosine kinase,selectively inhibits EGFR T790M/L858 R,but has weaker inhibitory effects on the WT EGFR in vitro. JAK3 inhibitor Ⅵ also specifically reduced autophosphorylation of EGFR T790M/L858 R in NCI-H1975 cells upon EGF stimulation,but did not show the inhibitory effect on WT EGFR in A431 cells. Furthermore,JAK3 inhibitor Ⅵ suppressed the proliferationof NCI-H1975 cells,but showed limited inhibitory effects on the WT EGFR-expressing cell lines A431 and A549.A docking simulation between JAK3 inhibitor Ⅵ and the ATP-binding pocket of EGFR T790M/L858 R predicted a potential binding status with hydrogen bonds. Estimated binding energy of JAK3 inhibitor Ⅵ to EGFR T790M/L858 R was more stable than its binding energy to the WT EGFR. Amino acid sequence alignments revealed that the gatekeeper residues of JAK family kinases are methionine in WT,similar to EGFR T790 M,suggesting that TKIs for JAKs may also be effective for EGFR T790 M. CONCLUSION:Our findings demonstrate that JAK3 inhibitor Ⅵ is a gatekeeper mutant selective TKI and offer a strategy to search for new EGFR T790 M inhibitors.
文摘Staphylococcus aureus is pathogenic to humans with worldwide health care concern due to its ability to evade the immune system and develop resistance to multiple drugs. Reg family proteins are C-type lectins with antimicrobial properties. Bacterial aggregation through binding to microbial cell surface sugar and/or lipid moieties is key mechanism employed in the process. In the present study we have analysed the antimicrobial effect of human REG Iα on S. aureus. Aggregation of mid-log phase culture of S. aureus was observed in presence of purified recombinant REG Iα. Therefore REG Iα can be applied in eliminating S. aureus infections in humans.
文摘Riboflavin （Rf） receptors bind and translocate Rf and its phosphorylated forms （e.g. flavin mononucleotide, FMN） into cells where they mediate various cellular metabolic pathways. Previously, we showed that FMN-coated ultrasmall superparamagnetic iron oxide （FLUSPIO） nanoparticles are suitable for labeling metabolically active cancer and endothelial cells in vitro. In this study, we focused on the in vivo application of FLUSPIO using prostate cancer xenografts. Size, charge, and chemical composition of FLUSPIO were evaluated. We explored the in vitro specificity of FLUSPIO for its cellular receptors using magnetic resonance imaging （MRI） and Prussian blue staining. Competitive binding experiments were performed in vivo by injecting free FMN in excess. Bio-distribution of FLUSPIO was determined by estimating iron content in organs and tumors using a colorimetric assay. AFM analysis and zeta potential measurements revealed a particulate morphology approximately 20-40 nm in size and a negative zeta potential （-24.23±0.15 mV） in water. X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry data confirmed FMN present on the USPIO nanoparticle surface. FLUSPIO uptake in prostate cancer cells and human umbilical vein endothelial cells was significantly higher than that of control USPIO, while addition of excess of free FMN reduced accumulation. Similarly, in vivo MRI and histology showed specific FLUSPIO uptake by prostate cancer cells, tumor endothelial cells, and tumor-associated macrophages. Besides prominent tumor accumulation, FLUSPIO accumulated in the liver, spleen, lung, and skin. Hence, our data strengthen our hypothesis that targeting riboflavin receptors is an efficient approach to accumulate nanomedicines in tumors opening perspectives for the development of diagnostic and therapeutic systems.
基金supported by the Van Andel Institute(to K.M.)the National Science Foundation(NSF+6 种基金MCB-1922846 to K.M.)the Six Talent Peaks Project in Jiangsu Province(NY-035 to F.Z.)the Fok Ying Tong Education Foundation(161022 to F.Z.)the National Institutes of Health(grant R01 GM57795 to G.A.H.)the Chemical Sciences,Geosciences,and Biosciences Division,Basic Energy Sciences,Office of Science at the U.S.Department of Energy(grant DE–FG02–91ER20021 to G.A.H.for infrastructure support)the Michigan State University Plant Resilience Institute(for support of L.V.-C.)the Michigan AgBioResearch Project(grant MICL02278 to G.A.H.).
文摘Jasmonates(JAs)are plant hormones with crucial roles in development and stress resilience.They activate MYC transcription factors by mediating the proteolysis of MYC inhibitors called JAZ proteins.In the absence of JA,JAZ proteins bind and inhibit MYC through the assembly of MYC–JAZ–Novel Interactor of JAZ(NINJA)–TPL repressor complexes.However,JAZ and NINJA are predicted to be largely intrinsically unstructured,which has precluded their experimental structure determination.Through a combination of biochemical,mutational,and biophysical analyses and AlphaFold-derived ColabFold modeling,we characterized JAZ–JAZ and JAZ–NINJA interactions and generated models with detailed,high-confidence domain interfaces.We demonstrate that JAZ,NINJA,and MYC interface domains are dynamic in isolation and become stabilized in a stepwise order upon complex assembly.By contrast,most JAZ and NINJA regions outside of the interfaces remain highly dynamic and cannot be modeled in a single conformation.Our data indicate that the small JAZ Zinc finger expressed in Inflorescence Meristem(ZIM)motif mediates JAZ–JAZ and JAZ–NINJA interactions through separate surfaces,and our data further suggest that NINJA modulates JAZ dimerization.This study advances our understanding of JA signaling by providing insights into the dynamics,interactions,and structure of the JAZ–NINJA core of the JA repressor complex.
基金supported by grants from the National Natural Science Foundation of China(82160770,81960741)the Guizhou Provincial Natural Science Foundation(QKH-J-2020-1Z070)+2 种基金Outstanding Young Scientific and Technological Talents Project of Guizhou Province(20215639)Zunyi Science and Technology Talent Platform Carrier Construction Project(ZSKRPT20231)Scientific and Technological Innovation Talent Team of Guizhou Province(CXTD[2023]024).
文摘Wound healing is a complex and multistep biological process that involves the cooperation of various cell types.Programmed cell death,including apoptosis and necrotizing apoptosis,plays a crucial role in this process.Apoptosis,a controlled and orderly programmed cell death regulated by genes,helps eliminate unnecessary or abnormal cells and maintain internal environmental stability.It also regulates various cell functions and contributes to the development of many diseases.In wound healing,programmed cell death is essential for removing inflammatory cells and forming scars.On the other hand,necroptosis,another form of programmed cell death,has not been thoroughly investigated regarding its role in wound healing.This review explores the changes and apoptosis of specific cell groups during wound healing after an injury and delves into the potential underlying mechanisms.Furthermore,it briefly discusses the possible mechanisms linking wound inflammation and fibrosis to apoptosis in wound healing.By understanding the relationship between apoptosis and wound healing and investigating the molecular mechanisms involved in apoptosis regulation,new strategies for the clinical treatment of wound healing may be discovered.
基金M.G-G and U.E have received funding from the postdoctoral fellowship programme Beatriu de Pinós,funded by the Secretary of Universities and Research(Government of Catalonia)and by the Horizon 2020 programme of research and innovation of the European Union under the Marie Sklodowska-Curie grant agreement No 801370R.A.P is supported by the Spanish Ministry by the Ram´on y Cajal Program(RYC2018-025977-I)Additional financial support was provided by the Government of Catalonia(2017 SGR 708)and MINECO/FEDER project(RTI2018-096088-J-100).
文摘Bacterial infection of implanted scaffolds may have fatal consequences and,in combination with the emergence of multidrug bacterial resistance,the development of advanced antibacterial biomaterials and constructs is of great interest.Since decades ago,metals and their ions had been used to minimize bacterial infection risk and,more recently,metal-based nanomaterials,with improved antimicrobial properties,have been advocated as a novel and tunable alternative.A comprehensive review is provided on how metal ions and ion nanoparticles have the potential to decrease or eliminate unwanted bacteria.Antibacterial mechanisms such as oxidative stress induction,ion release and disruption of biomolecules are currently well accepted.However,the exact antimicrobial mechanisms of the discussed metal compounds remain poorly understood.The combination of different metal ions and surface decorations of nanoparticles will lead to synergistic effects and improved microbial killing,and allow to mitigate potential side effects to the host.Starting with a general overview of antibacterial mechanisms,we subsequently focus on specific metal ions such as silver,zinc,copper,iron and gold,and outline their distinct modes of action.Finally,we discuss the use of these metal ions and nanoparticles in tissue engineering to prevent implant failure.
基金supported by the National Institute of General Medical Sciences of the National Institutes of Health under award number R35GM118041.
文摘CYP3A5 is a cytochrome P450(CYP)enzyme that metabolizes drugs and contributes to drug resistance in cancer.However,it remains unclear whether CYP3A5 directly influences cancer progression.In this report,we demonstrate that CYP3A5 regulates glucose metabolism in pancreatic ductal adenocarcinoma.Multi-omics analysis showed that CYP3A5 knockdown re-sults in a decrease in various glucose-related metabolites through its effect on glucose trans-port.A mechanistic study revealed that CYP3A5 enriches the glucose transporter GLUT1 at the plasma membrane by restricting the translation of TXNIP,a negative regulator of GLUT1.Notably,CYP3A5-generated reactive oxygen species were proved to be responsible for atten-uating the AKT-4EBP1-TXNIP signaling pathway.CYP3A5 contributes to cell migration by maintaining high glucose uptake in pancreatic cancer.Taken together,our results,for the first time,reveal a role of CYP3A5 in glucose metabolism in pancreatic ductal adenocarcinoma and identify a novel mechanism that is a potential therapeutic target.
基金supported by the fund forthe Joint Project of Bejing(Bejing Municipal Commission of Education)the National Natural Science of China(Grant Number:61727813)。
文摘Precise and high-speed sorting of individual target cells from heterogeneous populations plays an imperative role in cell research.Although the conventional fluorescence-activated cell sorter(FACs)is capable of rapid and accurate cell sorting,it occupies a large volume of the instrument and inherently brings in aerosol generation as well as crosscontamination among samples.The sorting completed in a fully enclosed and disposable microfluidic chip has the potential to eliminate the above concerns.However,current microfluidic cell sorters are hindered by the high.complexities of the fabrication procedure and the off-chip setup.In this paper,a spark-cavitation-bubble-based fluorescence-activated cell sorter is developed to perform fast and accurate sorting in a microfluidic chip.It features a simple structure and an easy operation.This microfluidic sorter comprises a positive electrode of platinum and a negative electrode of tungsten,which are placed on the side of the main channel.By applying a high-voltage discharge on the pair of electrodes,a single spark cavitation bubble is created to deflect the target particle into the downstream collection channel.The sorter has a short switching time of 150μs and a long lifespan of more than 100 million workable actions.In addition,a novel control strategy is proposed to dynamically adjust the discharge time to stabilize the size of the cavitation bubble for continuous sorting.The dynamic control of continuously triggering the sorter,the optimal delay time between fluorescence detection and cell sorting,and a theoretical model to predict the ideal sorting recovery and purity are studied to improve and evaluate the sorter performance.The experiments demonstrate that the sorting rate of target particles achieves 1200 eps,the total analysis throughput is up to 10,000 eps,the particles sorted at 4000 eps exhibit a purity greater than 80%and a recovery rate greater than 90%,and the sorting effect on the viability of HelLa cells is negligible.
基金supported by the National Key R&D Program of China(No.2022YFB4702902)National Natural Science Foundation of China(Nos.61831014,62275023,and 32021002)+2 种基金Beijing Municipal Natural Science Foundation(No.4232077)Overseas Expertise Introduction Project for Discipline Innovation(No.B18005)STI2030-Major Projects(No.2022ZD0212000).
文摘Optical imaging techniques provide low-cost,non-radiative images with high spatiotemporal resolution,making them advantageous for long-term dynamic observation of blood perfusion in stroke research and other brain studies compared to non-optical methods.However,high-resolution imaging in optical microscopy fundamentally requires a tight optical focus,and thus a limited depth of field(DOF).Consequently,large-scale,non-stitched,high-resolution images of curved surfaces,like brains,are difficult to acquire without z-axis scanning.To overcome this limitation,we developed a needle-shaped beam optical coherence tomography angiography(NB-OCTA)system,and for the first time,achieved a volumetric resolution of less than 8μm in a non-stitched volume space of 6.4 mm×4 mm×620μm in vivo.This system captures the distribution of blood vessels at 3.4-times larger depths than normal OCTA equipped with a Gaussian beam(GB-OCTA).We then employed NB-OCTA to perform long-term observation of cortical blood perfusion after stroke in vivo,and quantitatively analyzed the vessel area density(VAD)and the diameters of representative vessels in different regions over 10 days,revealing different spatiotemporal dynamics in the acute,sub-acute and chronic phase of post-ischemic revascularization.Benefiting from our NB-OCTA,we revealed that the recovery process is not only the result of spontaneous reperfusion,but also the formation of new vessels.This study provides visual and mechanistic insights into strokes and helps to deepen our understanding of the spontaneous response of brain after stroke.
基金support to the Xie laboratory from the National Institutes of Health and the National Institute on Drug Abuse(R01 DA052329).
文摘Proteins play a critical role in biology and biopharma due to their specificity and minimal side effects.Predicting the effects of mutations on protein stability is vital but experimentally challenging.Deep learning offers an efficient solution to this problem.In the present work,we introduced ProstaNet,a deep learning framework that predicts stability changes resulting from single-and multiple-point mutations using geometric vector perceptrons-graph neural network for 3-dimensional feature processing.For training ProstaNet,we meticulously crafted ProstaDB,a comprehensive and pristine thermodynamics repository,including 3,784 single-point mutations and 1,642 multiple-point mutations.We also created thermodynamic looping for enlarging the limited data size of multiple-point mutation and applied an innovative clustering method to generate a standard testing set of multiple-point mutation.Besides,we identified residue scoring as the most important encoding method in protein properties prediction.With these innovations,ProstaNet accurately predicts thermostability changes for both single-point and multiple-point mutations without showing any bias.ProstaNet achieves an accuracy of 0.75,outperforming existing methods for single-point mutation prediction,including ThermoMPNN(0.63),PoPMuSiC^(sym)(0.66),MUPRO(0.52),and FoldX(0.71).ProstaNet also achieves a 1.3-fold increase in accuracy compared to FoldX for multiple-point mutation predictions.Validated by experiment,4 out of 5 single-point mutation predictions(80%)and all multiple-point mutation predictions(100%)for HuJ3 mutants were accurate,demonstrating the potential benefits of ProstaNet for protein engineering and drug development.
