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Title: Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis

Abstract

The chemical cross-linking/mass spectrometry (MS) approach is gaining increasing importance as an alternative method for studying protein conformation and for deciphering protein interaction networks. This study is part of our ongoing efforts to develop innovative cross-linking principles for a facile and efficient assignment of cross-linked products. We evaluate two homobifunctional, amine-reactive, and MS-cleavable cross-linkers regarding their potential for automated analysis of cross-linked products. We introduce the bromine phenylurea (BrPU) linker that possesses a unique structure yielding a distinctive fragmentation pattern on collisional activation. Moreover, BrPU delivers the characteristic bromine isotope pattern and mass defect for all cross-linker-decorated fragments. We compare the fragmentation behavior of the BrPU linker with that of our previously described MS-cleavable TEMPO-Bz linker (which consists of a 2,2,6,6-tetramethylpiperidine-1-oxy moiety connected to a benzyl group) that was developed to perform free-radical-initiated peptide sequencing. Comparative collisional activation experiments (collision-induced dissociation and higher-energy collision-induced dissociation) with both cross-linkers were conducted in negative electrospray ionization mode with an Orbitrap Fusion mass spectrometer using five model peptides. As hypothesized in a previous study, the presence of a cross-linked N-terminal aspartic acid residue seems to be the prerequisite for the loss of an intact peptide from the cross-linked products. As the BrPU linkermore » combines a characteristic mass shift with an isotope signature, it presents a more favorable combination for automated assignment of cross-linked products compared with the TEMPO-Bz linker. .« less

Authors:
 [1];  [2];  [3];  [1]
  1. Martin Luther University Halle-Wittenberg, Institute of Pharmacy (Germany)
  2. Eurofins Umwelt West GmbH (Germany)
  3. University of Cologne, Department of Chemistry (Germany)
Publication Date:
OSTI Identifier:
22776883
Resource Type:
Journal Article
Journal Name:
Journal of the American Society for Mass Spectrometry
Additional Journal Information:
Journal Volume: 28; Journal Issue: 10; Other Information: Copyright (c) 2017 American Society for Mass Spectrometry; http://www.springer-ny.com; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 1044-0305
Country of Publication:
United States
Language:
English
Subject:
46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND TECHNOLOGY; COLLISIONS; COMPARATIVE EVALUATIONS; CROSS-LINKING; FRAGMENTATION; IONIZATION; MASS DEFECT; MASS SPECTROMETERS; MASS SPECTROSCOPY; PEPTIDES

Citation Formats

Hage, Christoph, Falvo, Francesco, Schäfer, Mathias, and Sinz, Andrea. Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis. United States: N. p., 2017. Web. doi:10.1007/S13361-017-1712-1.
Hage, Christoph, Falvo, Francesco, Schäfer, Mathias, & Sinz, Andrea. Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis. United States. https://doi.org/10.1007/S13361-017-1712-1
Hage, Christoph, Falvo, Francesco, Schäfer, Mathias, and Sinz, Andrea. Sun . "Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis". United States. https://doi.org/10.1007/S13361-017-1712-1.
@article{osti_22776883,
title = {Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis},
author = {Hage, Christoph and Falvo, Francesco and Schäfer, Mathias and Sinz, Andrea},
abstractNote = {The chemical cross-linking/mass spectrometry (MS) approach is gaining increasing importance as an alternative method for studying protein conformation and for deciphering protein interaction networks. This study is part of our ongoing efforts to develop innovative cross-linking principles for a facile and efficient assignment of cross-linked products. We evaluate two homobifunctional, amine-reactive, and MS-cleavable cross-linkers regarding their potential for automated analysis of cross-linked products. We introduce the bromine phenylurea (BrPU) linker that possesses a unique structure yielding a distinctive fragmentation pattern on collisional activation. Moreover, BrPU delivers the characteristic bromine isotope pattern and mass defect for all cross-linker-decorated fragments. We compare the fragmentation behavior of the BrPU linker with that of our previously described MS-cleavable TEMPO-Bz linker (which consists of a 2,2,6,6-tetramethylpiperidine-1-oxy moiety connected to a benzyl group) that was developed to perform free-radical-initiated peptide sequencing. Comparative collisional activation experiments (collision-induced dissociation and higher-energy collision-induced dissociation) with both cross-linkers were conducted in negative electrospray ionization mode with an Orbitrap Fusion mass spectrometer using five model peptides. As hypothesized in a previous study, the presence of a cross-linked N-terminal aspartic acid residue seems to be the prerequisite for the loss of an intact peptide from the cross-linked products. As the BrPU linker combines a characteristic mass shift with an isotope signature, it presents a more favorable combination for automated assignment of cross-linked products compared with the TEMPO-Bz linker. .},
doi = {10.1007/S13361-017-1712-1},
url = {https://www.osti.gov/biblio/22776883}, journal = {Journal of the American Society for Mass Spectrometry},
issn = {1044-0305},
number = 10,
volume = 28,
place = {United States},
year = {2017},
month = {10}
}