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Type: Artigo de periódico
Title: Chemical cross-linking with a diazirine photoactivatable cross-linker investigated by MALDI- and ESI-MS/MS
Author: Gomes, AF
Gozzo, FC
Abstract: Crystallography and nuclear magnetic resonance are well-established methods to study protein tertiary structure and interactions. Despite their usefulness, such methods are not applicable to many protein systems. Chemical cross-linking of proteins coupled with mass spectrometry allows low-resolution characterization of proteins and protein complexes based on measuring distance constraints from cross-links. In this work, we have investigated cross-linking by means of a heterobifunctional cross-linker containing a traditional N-hydroxysuccinimide (NHS) ester and a UV photoactivatable diazirine group. Activation of the diazirine group yields a highly reactive carbene species, with potential to increase the number of cross-links compared with homobifunctional, NHS-based cross-linkers. Cross-linking reactions were performed on model systems such as synthetic peptides and equine myoglobin. After reduction of the disulfide bond, the formation of intra- and intermolecular cross-links was identified and the peptides modified with both NHS and diazirine moieties characterized. Fragmentation of these modified peptides reveals the presence of a marker ion for intramolecular cross-links, which facilitates identification. Copyright (c) 2010 John Wiley & Sons, Ltd.
Subject: cross-linking
mass spectrometry, diazirine
Country: Inglaterra
Editor: John Wiley & Sons Ltd
Citation: Journal Of Mass Spectrometry. John Wiley & Sons Ltd, v. 45, n. 8, n. 892, n. 899, 2010.
Rights: fechado
Identifier DOI: 10.1002/jms.1776
Date Issue: 2010
Appears in Collections:Unicamp - Artigos e Outros Documentos

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