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Anal Chem ; 71(15): 3226-30, 1999 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-10450164

RESUMO

Characteristic ions in the MALDI TOF mass spectra from bacterial cells have been associated with four known proteins. The proteins, observed both from cells and in filtered cellular suspensions, were isolated by HPLC and identified on the basis of their mass spectra and their partial amino acid sequence, determined using the Edman method (10-15 residues). The acid resistance proteins HdeA and HdeB give rise to ions near m/z 9735 and 9060 in MALDI TOF mass spectra from cells and from extracts of both Escherichia coli 1090 and Shigella flexneri PHS-1059. However, the proteins associated with proteolytic cleavage by the peptidase Lep, rather than the precursor proteins, were observed, both using cells and from cellular extracts. A cold-shock protein, CspA, was associated with the ion near m/z 7643 from Pseudomonas aeruginosa. Similarly, a cold-acclimation protein, CapB, was identified as the source of the ion near m/z 7684 in P. putida. This last protein was homologous with a known CapB from P. fragi. While these experiments involved the detection of known or homologous proteins from typical bacteria, this same approach could also be applied to the detection of unique proteins or biomarker proteins associated with other bacteria of public health significance.


Assuntos
Proteínas de Bactérias/análise , Proteínas de Escherichia coli , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Sequência de Aminoácidos , Proteínas de Bactérias/isolamento & purificação , Proteínas de Transporte/análise , Proteínas de Transporte/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Temperatura Baixa , Escherichia coli/química , Proteínas de Choque Térmico/análise , Proteínas de Choque Térmico/isolamento & purificação , Dados de Sequência Molecular , Pseudomonas aeruginosa/química , Homologia de Sequência de Aminoácidos , Shigella flexneri/química
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