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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.)

A novel mass spectrometric strategy ``BEMAP{''} reveals Extensive O-linked protein glycosylation in Enterotoxigenic Escherichia coli

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Autor(es):
Boysen, Anders ; Palmisano, Giuseppe ; Krogh, Thoger Jensen ; Duggin, Iain G. ; Larsen, Martin R. ; Moller-Jensen, Jakob
Número total de Autores: 6
Tipo de documento: Artigo Científico
Fonte: SCIENTIFIC REPORTS; v. 6, AUG 26 2016.
Citações Web of Science: 6
Resumo

The attachment of sugars to proteins via side-chain oxygen atoms (O-linked glycosylation) is seen in all three domains of life. However, a lack of widely-applicable analytical tools has restricted the study of this process, particularly in bacteria. In E. coli, only four O-linked glycoproteins have previously been characterized. Here we present a glycoproteomics technique, termed BEMAP, which is based on the beta-elimination of O-linked glycans followed by Michael-addition of a phosphonic acid derivative, and subsequent titanium dioxide enrichment. This strategy allows site-specific mass-spectrometric identification of proteins with O-linked glycan modifications in a complex biological sample. Using BEMAP we identified cell surface-associated and membrane vesicle glycoproteins from Enterotoxigenic E. coli (ETEC) and non-pathogenic E. coli K-12. We identified 618 glycosylated Serine and Threonine residues mapping to 140 proteins in ETEC, including several known virulence factors, and 34 in E. coli K-12. The two strains had 32 glycoproteins in common. Remarkably, the majority of the ETEC glycoproteins were conserved in both strains but nevertheless were only glycosylated in the pathogen. Therefore, bacterial O-linked glycosylation is much more extensive than previously thought, and is especially important to the pathogen. (AU)

Processo FAPESP: 14/06863-3 - Modificações pós-traducionais para o diagnóstico de câncer e doenças parasitárias: abordagens metodológicas e implicações biológicas
Beneficiário:Giuseppe Palmisano
Linha de fomento: Auxílio à Pesquisa - Apoio a Jovens Pesquisadores