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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Chemical cross-linking with a diazirine photoactivatable cross-linker investigated by MALDI- and ESI-MS/MS

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Autor(es):
Gomes, Alexandre F. ; Gozzo, Fabio C.
Número total de Autores: 2
Tipo de documento: Artigo Científico
Fonte: Journal of Mass Spectrometry; v. 45, n. 8, p. 892-899, AUG 2010.
Citações Web of Science: 17
Resumo

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. (AU)

Processo FAPESP: 08/57805-2 - Instituto de Bioanalítica
Beneficiário:Lauro Tatsuo Kubota
Linha de fomento: Auxílio à Pesquisa - Temático
Processo FAPESP: 04/14846-0 - Rede de proteoma do estado de São Paulo
Beneficiário:Fabio Cesar Gozzo
Linha de fomento: Auxílio à Pesquisa - Programa GENOMA