Fragmentation behavior of a thiourea-based reagent for protein structure analysis by collision-induced dissociative chemical cross-linking
Article first published online: 7 JUL 2010
Copyright © 2010 John Wiley & Sons, Ltd.
Journal of Mass Spectrometry
Volume 45, Issue 8, pages 880–891, August 2010
How to Cite
Müller, M. Q., Dreiocker, F., Ihling, C. H., Schäfer, M. and Sinz, A. (2010), Fragmentation behavior of a thiourea-based reagent for protein structure analysis by collision-induced dissociative chemical cross-linking. J. Mass Spectrom., 45: 880–891. doi: 10.1002/jms.1775
- Issue published online: 23 AUG 2010
- Article first published online: 7 JUL 2010
- Manuscript Accepted: 27 MAY 2010
- Manuscript Received: 12 APR 2010
- DFG Graduiertenkolleg 1026 ‘Conformational Transitions in Macromolecular Interactions’
- chemical cross-linking;
- protein structure analysis;
The fragmentation behavior of a novel thiourea-based cross-linker molecule specifically designed for collision-induced dissociation (CID) MS/MS experiments is described. The development of this cross-linker is part of our ongoing efforts to synthesize novel reagents, which create either characteristic fragment ions or indicative constant neutral losses (CNLs) during tandem mass spectrometry allowing a selective and sensitive analysis of cross-linked products. The new derivatizing reagent for chemical cross-linking solely contains a thiourea moiety that is flanked by two amine-reactive N-hydroxy succinimide (NHS) ester moieties for reaction with lysines or free N-termini in proteins. The new reagent offers simple synthetic access and easy structural variation of either length or functionalities at both ends. The thiourea moiety exhibits specifically tailored CID fragmentation capabilities—a characteristic CNL of 85 u—ensuring a reliable detection of derivatized peptides by both electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) tandem mass spectrometry and as such possesses a versatile applicability for chemical cross-linking studies. A detailed examination of the CID behavior of the presented thiourea-based reagent reveals that slight structural variations of the reagent will be necessary to ensure its comprehensive and efficient application for chemical cross-linking of proteins. Copyright © 2010 John Wiley & Sons, Ltd.