期刊
JOURNAL OF PROTEOME RESEARCH
卷 8, 期 10, 页码 4564-4578出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr9004597
关键词
comparative proteomics; iTRAQ; two-dimensional gel electrophoresis; mass spectrometry; Leptospira; virulence factors; pathogenesis
资金
- Natural Sciences and Engineering Research Council of Canada [327186]
- British Columbia Proteomics Network
- Canada Foundation for Innovation
- British Columbia Knowledge Development Fund
- NHMRC Q Martin Fellow
- Canada Research Chair in Molecular Pathogenesis
- Michael Smith Foundation for Health Research Scholar
Comparative global proteome analyses were performed on Leptospira interrogans serovar Copenhageni grown under conventional in vitro conditions and those mimicking in vivo conditions (iron limitation and serum presence). Proteomic analyses were conducted using iTRAQ and LC-ESI-tandem mass spectrometry complemented with two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry. A total of 563 proteins were identified in this study. Altered expression of 65 proteins, including upregulation of the L. interrogans virulence factor Loa22 and 5 novel proteins with homology to virulence factors found in other pathogens, was observed between the comparative conditions. Immunoblot analyses confirmed upregulation of 5 of the known or putative virulence factors in L. interrogans exposed to the in vivo-like environmental conditions. Further, ELISA analyses using serum from patients with leptospirosis and immunofluorescence studies performed on liver sections derived from L. interrogans-infected hamsters verified expression of all but one of the identified proteins during infection. These studies, which represent the first documented comparative global proteome analysis of Leptospira, demonstrated proteome alterations under conditions that mimic in vivo infection and allowed for the identification of novel putative L. interrogans virulence factors.
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