Hage, Christoph, Ihling, Christian H., Goetze, Michael, Schaefer, Mathias ORCID: 0000-0002-5943-4335 and Sinz, Andrea (2017). Dissociation Behavior of a TEMPO-Active Ester Cross-Linker for Peptide Structure Analysis by Free Radical Initiated Peptide Sequencing (FRIPS) in Negative ESI-MS. J. Am. Soc. Mass Spectrom., 28 (1). S. 56 - 69. NEW YORK: SPRINGER. ISSN 1879-1123

Full text not available from this repository.

Abstract

We have synthesized a homobifunctional amine-reactive cross-linking reagent, containing a TEMPO (2,2,6,6-tetramethylpiperidine-1-oxy) and a benzyl group (Bz), termed TEMPO-Bz-linker, to derive three-dimensional structural information of proteins. The aim for designing this novel cross-linker was to facilitate the mass spectrometric analysis of cross-linked products by free radical initiated peptide sequencing (FRIPS). In an initial study, we had investigated the fragmentation behavior of TEMPO-Bz-derivatized peptides upon collision activation in (+)-electrospray ionization collision-induced dissociation tandem mass spectrometry (ESI-CID-MS/MS) experiments. In addition to the homolytic NO-C bond cleavage FRIPS pathway delivering the desired odd-electron product ions, an alternative heterolytic NO-C bond cleavage, resulting in even-electron product ions mechanism was found to be relevant. The latter fragmentation route clearly depends on the protonation of the TEMPO-Bz-moiety itself, which motivated us to conduct (-)-ESI-MS, CID-MS/MS, and MS3 experiments of TEMPO-Bz-cross-linked peptides to further clarify the fragmentation behavior of TEMPO-Bz-peptide molecular ions. We show that the TEMPO-Bz-linker is highly beneficial for conducting FRIPS in negative ionization mode as the desired homolytic cleavage of the NO-C bond is the major fragmentation pathway. Based on characteristic fragments, the isomeric amino acids leucine and isoleucine could be discriminated. Interestingly, we observed pronounced amino acid side chain losses in cross-linked peptides if the cross-linked peptides contain a high number of acidic amino acids.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Hage, ChristophUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ihling, Christian H.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Goetze, MichaelUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schaefer, MathiasUNSPECIFIEDorcid.org/0000-0002-5943-4335UNSPECIFIED
Sinz, AndreaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-248042
DOI: 10.1007/s13361-016-1426-9
Journal or Publication Title: J. Am. Soc. Mass Spectrom.
Volume: 28
Number: 1
Page Range: S. 56 - 69
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:
KeywordsLanguage
PROTEIN-PROTEIN INTERACTIONS; MASS-SPECTROMETRY; AUTOMATED ASSIGNMENT; LINKING; IDENTIFICATION; FRAGMENTATION; REAGENT; CHEMISTRY; RESIDUES; DYNAMICSMultiple languages
Biochemical Research Methods; Chemistry, Analytical; Chemistry, Physical; SpectroscopyMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/24804

Downloads

Downloads per month over past year

Altmetric

Export

Actions (login required)

View Item View Item