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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, Alexandre F
Gozzo, Fabio C
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.
Subject: Animals
Chromatography, Liquid
Cross-linking Reagents
Mass Spectrometry
Citation: Journal Of Mass Spectrometry : Jms. v. 45, n. 8, p. 892-9, 2010-Aug.
Rights: fechado
Identifier DOI: 10.1002/jms.1776
Date Issue: 2010
Appears in Collections:Unicamp - Artigos e Outros Documentos

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