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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 fragmentation photoactivatable |
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 |
Files in This Item:
File | Description | Size | Format | |
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WOS000281569100005.pdf | 315.61 kB | Adobe PDF | View/Open |
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