Total synthesis of TRADD death domain with arginine N-GlcNAcylation by hydrazide-based native chemical ligation  

作　　者：Ye Wu Yulei Li Wei Cong Yan Zou Xiang Li Honggang Hu 

机构地区：[1]Institute of Translational Medicine,Shanghai University,Shanghai 200436,China [2]School of Pharmacy,Second Military Medical University,Shanghai 200433,China [3]Key Laboratory of Marine Drugs,Ministry of Education,School of Medicine and Pharmacy,Ocean University of China,Qingdao 266003,China

出　　处：《Chinese Chemical Letters》2020年第1期107-110,共4页中国化学快报（英文版）

基　　金：the National Natural Science Foundation of China (Nos.91849129,21807112);PLA Youth Medical Science and Technology Youth Development Program (No.16QNP086);Foundation of Second Military Medical University (No.2016JS11)

摘　　要：TNFR1-associated death domain protein(TRADD)with arginine N-GlcNAcylation is a novel and structurally unique posttranslational modification(PTM)glycoprotein that blocks the formation of death-inducing signaling complex(DISC),orchestrating host nuclear factorκB(NF-κB)signaling in entero-pathogenic Escherichia coli(EPEC)-infected cells.This particular glycosylated modification plays an extremely vital role for the effective colonization and pathogenesis of pathogens in the gut.Herein we describe the total synthesis of TRADD death domain(residues 195-312)with arginine235 NGlcNAcylation(Arg-GIcNAc TRADD(195-312)).Two longish peptidyl fragments of the wild-type primary sequence were obtained by robust,microwave-assisted,highly efficient,solid-phase peptide synthesis(SPPS),the N-GlcNAcylated sector was built by total synthesis and attached specifically to resinbound peptide with an unprotected ornithine residue via silver-promoted on-resin guanidinylation,ArgGlcNAc TRADD(195-312)was constructed by hydrazide-based native chemical ligation(NCL).The facile synthetic strategy is expected to be generally applicable for the rapid synthesis of other proteins with Arg-GIcNAc modification and to pave the way for the related chemically biological study.TNFR1-associated death domain protein(TRADD) with arginine N-GlcNAcylation is a novel and structurally unique posttranslational modification(PTM) glycoprotein that blocks the formation of death-inducing signaling complex(DISC),orchestrating host nuclear factor κB(NF-κB) signaling in entero-pathogenic Escherichia coli(EPEC)-infected cells.This particular glycosylated modification plays an extremely vital role for the effective colonization and pathogenesis of pathogens in the gut.Herein we describe the total synthesis of TRADD death domain(residues 195-312) with arginine235 NGlcNAcylation(Arg-GIcNAc TRADD(195-312)).Two longish peptidyl fragments of the wild-type primary sequence were obtained by robust,microwave-assisted,highly efficient,solid-phase peptide synthesis(SPPS),the N-GlcNAcylated sector was built by total synthesis and attached specifically to resinbound peptide with an unprotected ornithine residue via silver-promoted on-resin guanidinylation,ArgGlcNAc TRADD(195-312) was constructed by hydrazide-based native chemical ligation(NCL).The facile synthetic strategy is expected to be generally applicable for the rapid synthesis of other proteins with Arg-GIcNAc modification and to pave the way for the related chemically biological study.

关 键 词：TRADD death domain Posttranslational modification Protein chemical synthesis Solid-phase peptide synthesis Native chemical ligation Peptide hydrazide 

分 类 号：TQ931[化学工程]