Universität zu Köln

Hage, Christoph, Falvo, Francesco, Schaefer, Mathias ORCID: 0000-0002-5943-4335 and Sinz, Andrea (2017). Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis. J. Am. Soc. Mass Spectrom., 28 (10). S. 2022 - 2039. WASHINGTON: AMER CHEMICAL SOC. ISSN 1879-1123

Full text not available from this repository.

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 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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Hage, Christoph UNSPECIFIED UNSPECIFIED UNSPECIFIED Falvo, Francesco UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaefer, Mathias UNSPECIFIED orcid.org/0000-0002-5943-4335 UNSPECIFIED Sinz, Andrea UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-216795
DOI:	10.1007/s13361-017-1712-1
Journal or Publication Title:	J. Am. Soc. Mass Spectrom.
Volume:	28
Number:	10
Page Range:	S. 2022 - 2039
Date:	2017
Publisher:	AMER CHEMICAL SOC
Place of Publication:	WASHINGTON
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; SEQUENCING FRIPS; LINKING REAGENTS; DISSOCIATION BEHAVIOR; IDENTIFICATION; FRAGMENTATION; ION; ENRICHMENT Multiple languages Biochemical Research Methods; Chemistry, Analytical; Chemistry, Physical; Spectroscopy Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/21679