Universität zu Köln

Iacobucci, Claudio ORCID: 0000-0001-9592-3606, Hage, Christoph, Schaefer, Mathias ORCID: 0000-0002-5943-4335 and Sinz, Andrea (2017). A Novel MS-Cleavable Azo Cross-Linker for Peptide Structure Analysis by Free Radical Initiated Peptide Sequencing (FRIPS). J. Am. Soc. Mass Spectrom., 28 (10). S. 2039 - 2054. NEW YORK: SPRINGER. ISSN 1879-1123

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Abstract

The chemical cross-linking/mass spectrometry (MS) approach is a growing research field in structural proteomics that allows gaining insights into protein conformations. It relies on creating distance constraints between cross-linked amino acid side chains that can further be used to derive protein structures. Currently, the most urgent task for designing novel cross-linking principles is an unambiguous and automated assignment of the created cross-linked products. Here, we introduce the homobifunctional, amine-reactive, and water soluble cross-linker azobisimidoester (ABI) as a prototype of a novel class of cross-linkers. The ABI-linker possesses an innovative modular scaffold combining the benefits of collisional activation lability with open shell chemistry. This MS-cleavable cross-linker can be efficiently operated via free radical initiated peptide sequencing (FRIPS) in positive ionization mode. Our proof-of-principle study challenges the gas phase behavior of the ABI-linker for the three amino acids, lysine, leucine, and isoleucine, as well as the model peptide thymopentin. The isomeric amino acids leucine and isoleucine could be discriminated by their characteristic side chain fragments. Collisional activation experiments were conducted via positive electrospray ionization (ESI) on two Orbitrap mass spectrometers. The ABI-mediated formation of odd electron product ions in MS/MS and MS3 experiments was evaluated and compared with a previously described azo-based cross-linker. All cross-linked products were amenable to automated analysis by the MeroX software, underlining the future potential of the ABI-linker for structural proteomics studies.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Iacobucci, Claudio UNSPECIFIED orcid.org/0000-0001-9592-3606 UNSPECIFIED Hage, Christoph UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaefer, Mathias UNSPECIFIED orcid.org/0000-0002-5943-4335 UNSPECIFIED Sinz, Andrea UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-217055
DOI:	10.1007/s13361-017-1744-6
Journal or Publication Title:	J. Am. Soc. Mass Spectrom.
Volume:	28
Number:	10
Page Range:	S. 2039 - 2054
Date:	2017
Publisher:	SPRINGER
Place of Publication:	NEW YORK
ISSN:	1879-1123
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Chemistry > Institute of Organic Chemistry
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language PROTEIN-PROTEIN INTERACTIONS; MASS-SPECTROMETRY; ELECTRON-TRANSFER; LINKING REAGENTS; DISSOCIATION BEHAVIOR; AUTOMATED ASSIGNMENT; IDENTIFICATION; FRAGMENTATION; PROTEOMICS; ENRICHMENT Multiple languages Biochemical Research Methods; Chemistry, Analytical; Chemistry, Physical; Spectroscopy Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/21705