Universität zu Köln

Esch, Patrick, Fischer, Moritz, Heiles, Sven ORCID: 0000-0003-3779-8071 and Schaefer, Mathias ORCID: 0000-0002-5943-4335 (2019). Olefinic reagents tested for peptide derivatization with switchable properties: Stable upon collision induced dissociation and cleavable by in-source Paterno-Buchi reactions. J. Mass Spectrom., 54 (12). S. 976 - 987. HOBOKEN: WILEY. ISSN 1096-9888

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Abstract

This contribution is part of our ongoing efforts to develop innovative cross-linking (XL) reagents and protocols for facilitated peptide mixture analysis and efficient assignment of cross-linked peptide products. In this report, we combine in-source Paterno-Buchi (PB) photo-chemistry with a tandem mass spectrometry approach to selectively address the fragmentation of a tailor-made cross-linking reagent. The PB photochemistry, so far exclusively used for the identification of unsaturation sites in lipids and in lipidomics, is now introduced to the field of chemical cross-linking. Based on trans-3-hexenedioic acid, an olefinic homo bifunctional amine reactive XL reagent was designed and synthesized for this proof-of-principle study. Condensation products of the olefinic reagent with a set of exemplary peptides are used to test the feasibility of the concept. Benzophenone is photochemically reacted in the nano-electrospray ion source and forms oxetane PB reaction products. Subsequent CID-MS triggered retro-PB reaction of the respective isobaric oxetane molecular ions and delivers reliably and predictably two sets of characteristic fragment ions of the cross-linker. Based on these signature ion sets, a straightforward identification of covalently interconnected peptides in complex digests is proposed. Furthermore, CID-MSn experiments of the retro-PB reaction products deliver peptide backbone characteristic fragment ions. Additionally, the olefinic XL reagents exhibit a pronounced robustness upon CID-activation, without previous UV-excitation. These experiments document that a complete backbone fragmentation is possible, while the linker-moiety remains intact. This feature renders the new olefinic linkers switchable between a stable, noncleavable cross-linking mode and an in-source PB cleavable mode.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Esch, Patrick UNSPECIFIED UNSPECIFIED UNSPECIFIED Fischer, Moritz UNSPECIFIED UNSPECIFIED UNSPECIFIED Heiles, Sven UNSPECIFIED orcid.org/0000-0003-3779-8071 UNSPECIFIED Schaefer, Mathias UNSPECIFIED orcid.org/0000-0002-5943-4335 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-126956
DOI:	10.1002/jms.4474
Journal or Publication Title:	J. Mass Spectrom.
Volume:	54
Number:	12
Page Range:	S. 976 - 987
Date:	2019
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1096-9888
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 CHEMICAL CROSS-LINKING; MASS-SPECTROMETRY; ELECTROSPRAY-IONIZATION; THERMAL-DECOMPOSITION; UNSATURATED LIPIDS; PROTEIN; IDENTIFICATION; FRAGMENTATION; ESTERS; BOND Multiple languages Biochemical Research Methods; Chemistry, Analytical; Spectroscopy Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/12695