Multiple phytohormones,including gibberellin(GA),abscisic acid(ABA),and indole-3-acetic acid(IAA),regulate seed germination.In this study,a barley aldehyde oxidase 1(HvAO1)gene was identified,which is located near the...Multiple phytohormones,including gibberellin(GA),abscisic acid(ABA),and indole-3-acetic acid(IAA),regulate seed germination.In this study,a barley aldehyde oxidase 1(HvAO1)gene was identified,which is located near the SD2(seed dormancy 2)region at the telomeric end of chromosome 5H.A doubledhaploid population(AC Metcalfe/Baudin)was used to characterize HvAO1 and validated its association with seed germination and malting quality.Aldehyde oxidase is predicted to catalyse the oxidation of various aldehydes,such as indoleacetaldehyde and abscisic aldehyde,into IAA and ABA,which is the final step of IAA/ABA biogenesis.This process influences the final IAA/ABA concentration in the seed,affecting the seed dormancy.Sequence analysis revealed substantial variations in the HvAO1 promoter regions between AC Metcalfe and Baudin.The combining seed germination tests,genetic variation analysis,gene expression,and phytohormone measurements showed that Baudin,which displays strong seed dormancy,has a specific sequence variation in the promoter region of the HvAO1 gene.This variation is associated with a higher expression level of the HvAO1 gene and an increased level of ABA than those in AC Metcalfe,which shows weak dormancy and lacks this sequence variation.In addition to its strong effect on the SD2 gene,HvAO1 shows excellent potential to fine-tune malting quality and seed dormancy,as evidenced by genotyping with HvAO1-specific markers,dormancy phenotypes,and malting quality.Our findings provide a new strategy for introducing favourable HvAO1 alleles to achieve the desired level of seed dormancy and high malting quality in barley.展开更多
Chiral pollutants often pose significant differential environmental health risks.In this study,the biotransformation of chiral dinotefuran(DIN)and its enantioselective metabolic toxicity mechanisms have been systemica...Chiral pollutants often pose significant differential environmental health risks.In this study,the biotransformation of chiral dinotefuran(DIN)and its enantioselective metabolic toxicity mechanisms have been systemically investigated.Firstly,reversedphase chromatography-high resolution mass spectrometry was developed to quantify the content of DIN R/S chiral enantiomer with pg level sensitivity,revealing a lower elimination rate constant(K_(e))of S-DIN(0.730 h^(-1))than R-DIN(0.746 h^(-1)).Secondly,the interaction mechanism between DIN metabolism and important endogenous bioactive molecules,such as aldehyde oxidase(AOX)and neurotransmitters,was revealed.The DIN nitro-group was converted into a guanidine group by the reducing site of nearby flavin adenine dinucleotide(FAD)in AOX with the preferred higher affinity of S-configuration.Meanwhile,the endogenous tryptophan(Trp)aldehyde metabolic intermediate,5-hydroxyindoleacetaldehyde(5-HIAL),provides a persistent electron donor for DIN reduction via the oxidation-catalyzed site in AOX,resulting in remarkable up-regulation of monoamine neurotransmitters such as serotonin and dopamine.Thirdly,the higher level of neurotransmitters further mediated dysregulation of oxylipin homeostasis via the serotonergic pathway,where S-DIN exhibited more pronounced liver lipid damage and environmental health risk with the accumulated lipid biomarkers,oxidized triglyceride(OxTG)and oxidized sphingomyelin(OxSM).This study elucidates the AOX-mediated enantioselectivity metabolic pathway of DIN,providing a new analytical method for chiral pollutants and paves the way for their health risk assessments.展开更多
Aldehyde oxidase(AOX)is a molybdoenzyme that is primarily expressed in the liver and is involved in the metabolism of drugs and other xenobiotics.AOX-mediated metabolism can result in unexpected outcomes,such as the p...Aldehyde oxidase(AOX)is a molybdoenzyme that is primarily expressed in the liver and is involved in the metabolism of drugs and other xenobiotics.AOX-mediated metabolism can result in unexpected outcomes,such as the production of toxic metabolites and high metabolic clearance,which can lead to the clinical failure of novel therapeutic agents.Computational models can assist medicinal chemists in rapidly evaluating the AOX metabolic risk of compounds during the early phases of drug discovery and provide valuable clues for manipulating AOX-mediated metabolism liability.In this study,we developed a novel graph neural network called AOMP for predicting AOX-mediated metabolism.AOMP integrated the tasks of metabolic substrate/non-substrate classification and metabolic site prediction,while utilizing transfer learning from 13C nuclear magnetic resonance data to enhance its performance on both tasks.AOMP significantly outperformed the benchmark methods in both cross-validation and external testing.Using AOMP,we systematically assessed the AOX-mediated metabolism of common fragments in kinase inhibitors and successfully identified four new scaffolds with AOX metabolism liability,which were validated through in vitro experiments.Furthermore,for the convenience of the community,we established the first online service for AOX metabolism prediction based on AOMP,which is freely available at https://aomp.alphama.com.cn.展开更多
基金supported by the Engineering Research Center of Ecology and Agricultural Use of Wetland,Ministry of Education(KFT202302)the National Natural Science Foundation of China(32372052).
文摘Multiple phytohormones,including gibberellin(GA),abscisic acid(ABA),and indole-3-acetic acid(IAA),regulate seed germination.In this study,a barley aldehyde oxidase 1(HvAO1)gene was identified,which is located near the SD2(seed dormancy 2)region at the telomeric end of chromosome 5H.A doubledhaploid population(AC Metcalfe/Baudin)was used to characterize HvAO1 and validated its association with seed germination and malting quality.Aldehyde oxidase is predicted to catalyse the oxidation of various aldehydes,such as indoleacetaldehyde and abscisic aldehyde,into IAA and ABA,which is the final step of IAA/ABA biogenesis.This process influences the final IAA/ABA concentration in the seed,affecting the seed dormancy.Sequence analysis revealed substantial variations in the HvAO1 promoter regions between AC Metcalfe and Baudin.The combining seed germination tests,genetic variation analysis,gene expression,and phytohormone measurements showed that Baudin,which displays strong seed dormancy,has a specific sequence variation in the promoter region of the HvAO1 gene.This variation is associated with a higher expression level of the HvAO1 gene and an increased level of ABA than those in AC Metcalfe,which shows weak dormancy and lacks this sequence variation.In addition to its strong effect on the SD2 gene,HvAO1 shows excellent potential to fine-tune malting quality and seed dormancy,as evidenced by genotyping with HvAO1-specific markers,dormancy phenotypes,and malting quality.Our findings provide a new strategy for introducing favourable HvAO1 alleles to achieve the desired level of seed dormancy and high malting quality in barley.
基金supported by the National Natural Science Foundation of China(22276076,22306074,22361132536,22236002)the Fundamental Research Funds for the Central Universities(JUSRP622032)the Jiangsu Association for Science and Technology Youth Science and Technology Talent Support Project(TJ2021-049)。
文摘Chiral pollutants often pose significant differential environmental health risks.In this study,the biotransformation of chiral dinotefuran(DIN)and its enantioselective metabolic toxicity mechanisms have been systemically investigated.Firstly,reversedphase chromatography-high resolution mass spectrometry was developed to quantify the content of DIN R/S chiral enantiomer with pg level sensitivity,revealing a lower elimination rate constant(K_(e))of S-DIN(0.730 h^(-1))than R-DIN(0.746 h^(-1)).Secondly,the interaction mechanism between DIN metabolism and important endogenous bioactive molecules,such as aldehyde oxidase(AOX)and neurotransmitters,was revealed.The DIN nitro-group was converted into a guanidine group by the reducing site of nearby flavin adenine dinucleotide(FAD)in AOX with the preferred higher affinity of S-configuration.Meanwhile,the endogenous tryptophan(Trp)aldehyde metabolic intermediate,5-hydroxyindoleacetaldehyde(5-HIAL),provides a persistent electron donor for DIN reduction via the oxidation-catalyzed site in AOX,resulting in remarkable up-regulation of monoamine neurotransmitters such as serotonin and dopamine.Thirdly,the higher level of neurotransmitters further mediated dysregulation of oxylipin homeostasis via the serotonergic pathway,where S-DIN exhibited more pronounced liver lipid damage and environmental health risk with the accumulated lipid biomarkers,oxidized triglyceride(OxTG)and oxidized sphingomyelin(OxSM).This study elucidates the AOX-mediated enantioselectivity metabolic pathway of DIN,providing a new analytical method for chiral pollutants and paves the way for their health risk assessments.
基金supported by the National Natural Science Foundation of China(T2225002,82273855 to Mingyue Zheng)Lingang Laboratory(LG202102-01-02 to Mingyue Zheng)+1 种基金the National Key Research and Development Program of China(2022YFC3400504 to Mingyue Zheng)the open fund of state key laboratory of Pharmaceutical Biotechnology,Nanjing University,China(KF-202301 to Mingyue Zheng).
文摘Aldehyde oxidase(AOX)is a molybdoenzyme that is primarily expressed in the liver and is involved in the metabolism of drugs and other xenobiotics.AOX-mediated metabolism can result in unexpected outcomes,such as the production of toxic metabolites and high metabolic clearance,which can lead to the clinical failure of novel therapeutic agents.Computational models can assist medicinal chemists in rapidly evaluating the AOX metabolic risk of compounds during the early phases of drug discovery and provide valuable clues for manipulating AOX-mediated metabolism liability.In this study,we developed a novel graph neural network called AOMP for predicting AOX-mediated metabolism.AOMP integrated the tasks of metabolic substrate/non-substrate classification and metabolic site prediction,while utilizing transfer learning from 13C nuclear magnetic resonance data to enhance its performance on both tasks.AOMP significantly outperformed the benchmark methods in both cross-validation and external testing.Using AOMP,we systematically assessed the AOX-mediated metabolism of common fragments in kinase inhibitors and successfully identified four new scaffolds with AOX metabolism liability,which were validated through in vitro experiments.Furthermore,for the convenience of the community,we established the first online service for AOX metabolism prediction based on AOMP,which is freely available at https://aomp.alphama.com.cn.
