Chemical Cross-linking With A Diazirine Photoactivatable Cross-linker Investigated By Maldi- And Esi-ms/ms.

dc.creatorGomes, Alexandre F
dc.creatorGozzo, Fabio C
dc.date2010-Aug
dc.date2015-11-27T13:18:03Z
dc.date2015-11-27T13:18:03Z
dc.date.accessioned2018-03-29T01:11:16Z
dc.date.available2018-03-29T01:11:16Z
dc.identifierJournal Of Mass Spectrometry : Jms. v. 45, n. 8, p. 892-9, 2010-Aug.
dc.identifier1096-9888
dc.identifier10.1002/jms.1776
dc.identifierhttp://www.ncbi.nlm.nih.gov/pubmed/20635431
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/198950
dc.identifier20635431
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1299183
dc.descriptionCrystallography 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.
dc.description45
dc.description892-9
dc.languageeng
dc.relationJournal Of Mass Spectrometry : Jms
dc.relationJ Mass Spectrom
dc.rightsfechado
dc.rightsCopyright 2010 John Wiley & Sons, Ltd.
dc.sourcePubMed
dc.subjectAnimals
dc.subjectChromatography, Liquid
dc.subjectCross-linking Reagents
dc.subjectCysteine
dc.subjectDiazomethane
dc.subjectDisulfides
dc.subjectHorses
dc.subjectMass Spectrometry
dc.subjectMyoglobin
dc.subjectPeptides
dc.subjectPhotochemistry
dc.titleChemical Cross-linking With A Diazirine Photoactivatable Cross-linker Investigated By Maldi- And Esi-ms/ms.
dc.typeArtículos de revistas


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