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Infection and Immunity, August 2000, p. 4725-4735, Vol. 68, No. 8
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Antigenic and Sequence Diversity in Gonococcal Transferrin-Binding Protein A

Cynthia Nau Cornelissen,1,* James E. Anderson,2 Ian C. Boulton,1,dagger and P. Frederick Sparling2,3

Department of Microbiology and Immunology, School of Medicine, Virginia Commonwealth University, Richmond, Virginia 23298,1 and Departments of Medicine2 and Microbiology and Immunology,3 School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599

Received 10 March 2000/Returned for modification 21 April 2000/Accepted 12 May 2000

Neisseria gonorrhoeae is a gram-negative pathogen that is capable of satisfying its iron requirement with human iron-binding proteins such as transferrin and lactoferrin. Transferrin-iron utilization involves specific binding of human transferrin at the cell surface to what is believed to be a complex of two iron-regulated, transferrin-binding proteins, TbpA and TbpB. The genes encoding these proteins have been cloned and sequenced from a number of pathogenic, gram-negative bacteria. In the current study, we sequenced four additional tbpA genes from other N. gonorrhoeae strains to begin to assess the sequence diversity among gonococci. We compared these sequences to those from other pathogenic bacteria to identify conserved regions that might be important for the structure and function of these receptors. We generated polyclonal mouse sera against synthetic peptides deduced from the TbpA sequence from gonococcal strain FA19. Most of these synthetic peptides were predicted to correspond to surface-exposed regions of TbpA. We found that, while most reacted with denatured TbpA in Western blots, only one antipeptide serum reacted with native TbpA in the context of intact gonococci, consistent with surface exposure of the peptide to which this serum was raised. In addition, we evaluated a panel of gonococcal strains for antigenic diversity using these antipeptide sera.

* Corresponding author. Mailing address: P.O. Box 980678, Richmond, VA 23298-0678. Phone: (804) 225-4121. Fax: (804) 828-9946. E-mail: cncornel{at}hsc.vcu.edu.

dagger Present address: Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario, Canada.

Infection and Immunity, August 2000, p. 4725-4735, Vol. 68, No. 8
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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