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Infection and Immunity, December 2009, p. 5262-5271, Vol. 77, No. 12
0019-9567/09/$08.00+0     doi:10.1128/IAI.00911-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Proteomic Analysis of Rickettsia parkeri Strain Portsmouth{triangledown}

Walairat Pornwiroon,1 Apichai Bourchookarn,1,2 Christopher D. Paddock,3 and Kevin R. Macaluso1*

Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Skip Bertman Drive, SVM-3213, Baton Rouge, Louisiana 70803,1 Department of Technology and Industries, Faculty of Science and Technology, Prince of Songkla University, Pattani 94000, Thailand,2 Infectious Disease Pathology Branch, Mailstop G-32, Centers for Disease Control and Prevention, 1600 Clifton Rd., Atlanta, Georgia 303333

Received 11 August 2009/ Returned for modification 28 August 2009/ Accepted 15 September 2009

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, β-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.

* Corresponding author. Mailing address: Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Skip Bertman Drive, SVM-3213, Baton Rouge, LA 70803. Phone: (225) 578-9677. Fax: (225) 578-9701. E-mail: kmacaluso{at}vetmed.lsu.edu

{triangledown} Published ahead of print on 21 September 2009.

Editor: R. P. Morrison

Infection and Immunity, December 2009, p. 5262-5271, Vol. 77, No. 12
0019-9567/09/$08.00+0     doi:10.1128/IAI.00911-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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