Ynamide Coupling Reagent for the Chemical Cross-Linking of Proteins in Live Cells

Shengrong Li, Chengjun Zhu, Qian Zhao, Zhi Min Zhang, Pinghua Sun, Zhengqiu Li

Research output: Journal article publicationJournal articleAcademic researchpeer-review

6 Citations (Scopus)

Abstract

Chemical cross-linking of proteins coupled with mass spectrometry analysis (CXMS) is a powerful method for the study of protein structure and protein-protein interactions (PPIs). However, the chemical probes used in the CXMS are limited to bidentate reactive warheads, and the available zero-length cross-linkers are restricted to 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) and 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMTMM). To alleviate this issue, an efficient coupling reagent, sulfonyl ynamide, was developed as a new zero-length cross-linker that can connect high-abundance carboxyl residues (D/E) with lysine (K) to form amide bonds in the absence of any catalyst. Significant improvement in the cross-linking efficiency and specificity in comparison with traditional EDC/NHS was achieved with model proteins, which includes inter- and intramolecular conjugations. The cross-linked structures were validated by X-ray crystallography. Importantly, this coupling reagent can be successfully used to capture interacting proteins in the whole proteome and can be a useful reagent for probing potential protein-protein interactions in situ.

Original languageEnglish
JournalACS Chemical Biology
DOIs
Publication statusAccepted/In press - 2023

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Fingerprint

Dive into the research topics of 'Ynamide Coupling Reagent for the Chemical Cross-Linking of Proteins in Live Cells'. Together they form a unique fingerprint.

Cite this