Histidine(His)bears a uniquely electron-deficient imidazole side chain and plays essential roles in protein interactions and enzyme-catalyzed processes.Modification of His C_(2)position offers a useful method to fine-...Histidine(His)bears a uniquely electron-deficient imidazole side chain and plays essential roles in protein interactions and enzyme-catalyzed processes.Modification of His C_(2)position offers a useful method to fine-tune histidine residues of proteins for their structural and functional study.Due to the moderately nucleophilic imidazole group,the chemoselective modification of histidine in proteins remains particularly challenging.Herein,we report a highly efficient method for the semisynthesis of chitin-binding protein21(CBP21)bearing various groups at the C_(2)position of His28.A combination of modern radical-mediated C-H alkylation and recombinant protein engineering offers a powerful strategy to decipher His functions.展开更多
Validamycin has been widely used as a specific competitive inhibitor of tre-halase.In our previous research,validamycin significantly inhibited trehalase activity and chitin synthesis in Diaphorina citri,resulting in ...Validamycin has been widely used as a specific competitive inhibitor of tre-halase.In our previous research,validamycin significantly inhibited trehalase activity and chitin synthesis in Diaphorina citri,resulting in abnormal phenotypes.However,the mechanism of validamycin's action on D.citri remains unclear.Here,using a comparative transcriptome analysis,464 differentially expressed genes(DEGs)in D.citri were identified after validamycin treatment.A Gene Ontology enrichment analysis revealed that these DEGs were mainly involved in“small molecule process”,“structural molecule ac-tivity”and“transition metal ion binding”.DEGs involved in chitin metabolism,cuticle synthesis and insecticide detoxification were validated by reverse transcription quantitative polymerase chain reaction.The RNA interference of D.citri chitinase-like protein EN03 and D.citri cuticle protein 7 genes significantly affected D.citri molting.Moreover,the recombinant chitinase-like protein EN03 exhibited a chitin-binding property,and an antimicrobial activity against Bacillus subtilis.This study provides a first insight into the molecular changes in D.citri after exposure to validamycin and identifies two effective RNA interference targets for D.citri control.展开更多
基金supported by the National Science Fund for National Natural Science Foundation of China(grant nos.92253302,22225701,22077080 and 22477078)the Shanghai Pilot Program for Basic Research-Shanghai Jiao Tong University(grant no.21TQ1400210)+1 种基金the Interdisciplinary Program of Shanghai Jiao Tong University(grant no.20230102)the Shanghai Municipal Science and Technology Major Project and the Department of Human Resources,and Social Security of Sichuan Province,China.
文摘Histidine(His)bears a uniquely electron-deficient imidazole side chain and plays essential roles in protein interactions and enzyme-catalyzed processes.Modification of His C_(2)position offers a useful method to fine-tune histidine residues of proteins for their structural and functional study.Due to the moderately nucleophilic imidazole group,the chemoselective modification of histidine in proteins remains particularly challenging.Herein,we report a highly efficient method for the semisynthesis of chitin-binding protein21(CBP21)bearing various groups at the C_(2)position of His28.A combination of modern radical-mediated C-H alkylation and recombinant protein engineering offers a powerful strategy to decipher His functions.
基金supported by the Natural Science Foundation of Jiangxi Province(20202BAB215004)the National Natural Science Foundation of China(31960116)+5 种基金the Educational Commission of Jiangxi Province of China(GJJ180747)Key Research and Development Plan of Jiangxi Province(20192BBF60057,2017ZDD02005)Ministry of Science and Technology of China(2018YFD0201504)the earmarked fund for Jiangxi Agriculture Research System(JXARS-07)Gan-nan Normal University-level bidding project(20162B08)Graduate student innovation Fund Project(YC2019-S392).
文摘Validamycin has been widely used as a specific competitive inhibitor of tre-halase.In our previous research,validamycin significantly inhibited trehalase activity and chitin synthesis in Diaphorina citri,resulting in abnormal phenotypes.However,the mechanism of validamycin's action on D.citri remains unclear.Here,using a comparative transcriptome analysis,464 differentially expressed genes(DEGs)in D.citri were identified after validamycin treatment.A Gene Ontology enrichment analysis revealed that these DEGs were mainly involved in“small molecule process”,“structural molecule ac-tivity”and“transition metal ion binding”.DEGs involved in chitin metabolism,cuticle synthesis and insecticide detoxification were validated by reverse transcription quantitative polymerase chain reaction.The RNA interference of D.citri chitinase-like protein EN03 and D.citri cuticle protein 7 genes significantly affected D.citri molting.Moreover,the recombinant chitinase-like protein EN03 exhibited a chitin-binding property,and an antimicrobial activity against Bacillus subtilis.This study provides a first insight into the molecular changes in D.citri after exposure to validamycin and identifies two effective RNA interference targets for D.citri control.
