Analyzing Bacterial Pathogenesis at Level of Proteome

Cash, Phillip

doi:10.1002/0471973165.ch13

Cited by 1 publication

(1 citation statement)

References 88 publications

(132 reference statements)

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“…A membrane proteome is the entire complement of membrane proteins present in a cell at a given condition and time. Synthesis of proteins will be induced or repressed in the situation of growing on the artificial culture medium, especially for intracellular bacteria pathogens (Cash, 2006), and therefore, the actual number of proteins, which are the sort of proteins associated with adhesion and invasion observed on a 2-DE gel, is expected to be much lower. We established a 2-DE map of the outer membrane subproteome of B. henselae using the sodium carbonate extraction method and the lauryl sarcosine extraction method.…”

Section: Discussionmentioning

confidence: 99%

Proteomic and bioinformatic analysis of outer membrane proteins of the protobacterium Bartonella henselae (Bartonellaceae)

Dm¹,

Qy²,

Zhao³

et al. 2011

Genet. Mol. Res.

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ABSTRACT. Bartonella henselae, an infectious agent causing catscratch disease and vasculoproliferative disorders in humans, is a fastidious facultative intracellular pathogen. The outer membrane proteins of B. henselae are key molecules that play a primary role in host-cell interactions. We isolated B. henselae outer membrane proteins, using the ionic detergent N-lauroyl sarcosine sodium salt and sodium carbonate, purification by two-dimensional (2-D) gel electrophoresis, and protein identification using mass spectrometry. Treatment with buffers containing ASB-14 and ZWITTERGENT 3-10 increased solubilization of B. henselae proteins, particularly proteins with basic pI. Three hundred and sixty- eight spots were detected from the sarcosine-insoluble outer membrane fraction; 94 distinct protein species were identified from 176 spots. In the outer membrane fraction from carbonate incubation, 471 spots were calculated and 259 spots were identified, which included 139 protein entries. There were six outer membrane proteins in the sarcosine-insoluble outer membrane fraction compared with nine outer membrane proteins from samples subjected to carbonate incubation. We used bioinformatic analysis to identify 44 outer membrane proteins by prediction of their domains and tertiary structures and documented the potential virulence factors. We established the 2-D reference maps of the outer membrane subproteome of B. henselae using the two different extraction methods, which were partly complementary to each other. Sodium carbonate extraction isolated low-abundance and basic proteins better than the lauroyl sarcosine sodium salt extraction, which enriched high-abundance porins.

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Section: Discussionmentioning

confidence: 99%