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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.)

Collision-Induced Dissociation of Lys-Lys Intramolecular Crosslinked Peptides

Texto completo
Autor(es):
Iglesias, Amadeu H. [1, 2] ; Santos, Luiz F. A. [1, 2] ; Gozzo, Fabio C. [1, 2]
Número total de Autores: 3
Afiliação do(s) autor(es):
[1] Univ Estadual Campinas, Inst Chem, BR-13083970 Campinas, SP - Brazil
[2] Ctr Struct & Mol Biol, Brazilian Synchrotron Light Source, Campinas, SP - Brazil
Número total de Afiliações: 2
Tipo de documento: Artigo Científico
Fonte: JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY; v. 20, n. 4, p. 557-566, APR 2009.
Citações Web of Science: 16
Resumo

The use of chemical crosslinking is an attractive tool that presents many advantages in the application of mass spectrometry to structural biology. The correct assignment of crosslinked peptides, how-ever, is still. a challenge because of the lack of detailed fragmentation studies on resultant species. In this work, the fragmentation patterns of intramolecular crosslinked peptides with disuccinimidyl suberate (DSS) has been devised by using a set on versatile, model peptides that resemble species found in crosslinking experiments with proteins. These peptides contain an acetylated N-terminus followed by a random sequence of residues containing two lysine residues separated by an arginine. After the crosslinking reaction, controlled trypsin digestion yields both intra- and intermolecular crosslinked peptides. In the present study we analyzed the fragmentation of matrix-assisted laser desorption/ionization-generated peptides crosslinked with DSS in which both lysines are found in the same peptide. Fragmentation starts in the linear moiety of the peptide, yielding regular b and y ions. Once it reaches the cyclic portion of the molecule, fragmentation was observed to occur either at the following peptide bond or at the peptide crosslinker amide bond. If the peptide crosslinker bond is cleaved, it fragments as a regular modified peptide, in which the DSS backbone remains attached to the first lysine. This fragmentation pattern resembles the fragmentation of modified peptides and may be identified by common automated search engines using DSS as a modification. If, on the other hand, fragmentation happens at the peptide bond itself, rearrangement of the last crosslinked lysine is observed and a product ion containing the crosslinker backbone and lysine (m/z 222) is formed. The detailed identification of fragment ions can help the development of softwares devoted to the MS/MS data analysis of crosslinked peptides. (J Am Soc Mass Spectrom 2009, 20, 557-566) (C) 2009 Published by Elsevier Inc. on behalf of American Society for Mass Spectrometry (AU)

Processo FAPESP: 04/14846-0 - Rede de proteoma do estado de São Paulo
Beneficiário:Fabio Cesar Gozzo
Modalidade de apoio: Auxílio à Pesquisa - Programa GENOMA