Identification of di-Boron cleavage events at Dha residues on recombinant protein in e.coli lysate

Neurokinin-A (NKA) is only active when its C-terminus is amidated. B2(OH2) is thought to interact with Dha and cleave the peptide, leaving the amide on the C-terminus of the left-hand peptide. PeptideID MS/MS experiment was performed in e.coli lysate with overevpressed recombinant NKA-eH3 to prove diBoron activity in biologically relevant context. Overall analysis of the Dha generation and consumption on e.coli lysate was also performed.

神经激肽A（Neurokinin-A, NKA）仅在其C端发生酰胺化修饰后才具备生物活性。二硼水合物（B2(OH2)）被认为可与脱氢丙氨酸（Dehydroalanine, Dha）相互作用并切割肽链，使酰胺基保留在左侧肽段的C端。为验证二硼试剂在生物学相关环境中的活性，以过表达重组NKA-eH3的大肠杆菌（E. coli）裂解液为实验体系开展了PeptideID串联质谱（MS/MS）实验。同时还对大肠杆菌裂解液中脱氢丙氨酸的生成与消耗情况进行了全面分析。