A Membrane-Permeable and Immobilized Metal Affinity Chromatography (IMAC) Enrichable Cross-Linking Reagent to Advance In Vivo Cross-Linking Mass Spectrometry

被引：48

作者：

Jiang, Pin-Lian ^{[1
]}

Wang, Cong ^{[1
]}

Diehl, Anne ^{[1
]}

Viner, Rosa ^{[2
]}

Etienne, Chris ^{[3
]}

Nandhikonda, Premchendar ^{[3
]}

Foster, Leigh ^{[3
]}

Bomgarden, Ryan D. ^{[3
]}

Liu, Fan ^{[1
,4
]}

机构：

[1] Leibniz Forschungsinst Mol Pharmakol Forschungsve, Dept Struct Biol, Campus Berlin Buch Robert Roessle Str 10, D-13125 Berlin, Germany

[2] Thermo Fisher Sci, 355 River Oaks Pkwy, San Jose, CA 95134 USA

[3] Thermo Fisher Sci, 3747N Meridian Rd, Rockford, IL 61101 USA

[4] PCharite Univ Med Berlin, Charitepl 1, D-10117 Berlin, Germany

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2022年 / 61卷 / 12期

基金：

欧洲研究理事会;

关键词：

Cross-linking; Mass spectrometry; Protein-protein interactions; Protein structures; Proteomics; PROTEIN-PROTEIN INTERACTIONS;

D O I：

10.1002/anie.202113937

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Cross-linking mass spectrometry (XL-MS) is an attractive method for the proteome-wide characterization of protein structures and interactions. Currently, the depth of in vivo XL-MS studies is lagging behind the established applications to cell lysates, because cross-linking reagents that can penetrate intact cells and strategies to enrich cross-linked peptides lack efficiency. To tackle these limitations, we have developed a phosphonate-containing cross-linker, tBu-PhoX, that efficiently permeates various biological membranes and can be robustly enriched using routine immobilized metal ion affinity chromatography. We have established a tBu-PhoX-based in vivo XL-MS approach that enables cross-links in intact human cells to be identified in high numbers with substantially reduced analysis time. Collectively, the developed cross-linker and XL-MS approach pave the way for the comprehensive XL-MS characterization of living systems.

引用

页数：5

共 16 条

[1] Mass-spectrometric exploration of proteome structure and function [J].