Proteomic analysis of Rickettsia parkeri strain portsmouth.
Rickettsia parkeri, a recently recognized pathogen of human, is one of several Rickettsia spp. in the United States that causes a spotted fever rickettsiosis. To gain insights into its biology and pathogenesis, we applied the proteomics approach to establish a two-dimensional gel proteome reference map and combined this technique with cell surface biotinylation to identify surface-exposed proteins of a low-passage isolate of R. parkeri obtained from a patient. We identified 91 proteins by matrix-assisted laser desorption ionization-tandem time of flight mass spectrometry. Of these, 28 were characterized as surface proteins, including virulence-related proteins (e.g., outer membrane protein A [OmpA], OmpB, beta-peptide, and RickA). Two-dimensional immunoblotting with serum from the R. parkeri-infected index patient was utilized to identify the immunoreactive proteins as potential targets for diagnosis and vaccine development. In addition to the known rickettsial antigens, OmpA and OmpB, we identified translation initiation factor 2, cell division protein FtsZ, and cysteinyl-tRNA synthetase as immunoreactive proteins. The proteome map with corresponding cell surface protein analysis and antigen detection will facilitate a better understanding of the mechanisms of rickettsial pathogenesis.
Pornwiroon
Walairat
W
Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana 70803, USA .
Bourchookarn
Apichai
A
Paddock
Christopher D
CD
Macaluso
Kevin R
KR
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis* / Q000276:immunology; D002522:Chlorocebus aethiops; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D020543:Proteome / Q000032:analysis*; D012281:Rickettsia / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D012282:Rickettsia Infections / Q000276:immunology / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014709:Vero Cells
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; D020543:Proteome