Global proteome analysis of Leptospira interrogans.
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.
Eshghi
Azad
A
Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada.
Cullen
Paul A
PA
Cowen
Laura
L
Zuerner
Richard L
RL
Cameron
Caroline E
CE
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D002633:Chemotaxis; D003030:Coagulase / Q000378:metabolism; D006224:Cricetinae; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism; D005407:Flagella / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling; D007921:Leptospira interrogans / Q000378:metabolism*; D013058:Mass Spectrometry; D008647:Mesocricetus; D008856:Microscopy, Fluorescence; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D037521:Virulence Factors / Q000032:analysis / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D003030:Coagulase; D020543:Proteome; D011994:Recombinant Proteins; D037521:Virulence Factors; D006026:Glycoside Hydrolases