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Infection and Immunity, November 2000, p. 6189-6195, Vol. 68, No. 11
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Protective Glycosylphosphatidylinositol-Anchored Membrane Protein of Plasmodium yoelii Trophozoites and Merozoites Contains Two Epidermal Growth Factor-Like Domains

James M. Burns Jr.,* Carla C. Belk, and Patricia D. Dunn

Department of Microbiology, Meharry Medical College, Nashville, Tennessee 37208

Received 3 April 2000/Returned for modification 14 June 2000/Accepted 9 August 2000

Using sera from mice immunized and protected against Plasmodium yoelii malaria, we identified a novel blood-stage antigen gene, pypag-2. The 2.1-kb pypag-2 cDNA contains a single open reading frame that encodes a 409-amino-acid protein with a predicted molecular mass of 46.8 kDa. Unlike many characterized plasmodial antigens, blocks of tandemly repeated amino acids are lacking in the pypAg-2 protein sequence. Recombinant pypAg-2, comprising the full-length protein minus the predicted N-terminal signal and C-terminal anchor sequences, was produced and used to raise a high-titer polyclonal rabbit antiserum. This antiserum was used to identify and characterize the native protein through immunoblotting, immunoprecipitation and immunofluorescence assays. Consistent with the presence of a glycosylphosphatidylinositol anchor, pypAg-2 fractionated with the detergent phase of Triton X-114-solubilized proteins and could be metabolically labeled with [3H]palmitic acid. By immunofluorescence, pypAg-2 expression was localized to both the trophozoite and merozoite membranes. Similar to Plasmodium falciparum merozoite surface protein 1, pypAg-2 contains two C-terminal epidermal growth factor (EGF)-like domains. Most importantly, immunization with recombinant pypAg-2 protected mice against lethal P. yoelii malaria. Thus, pypAg-2 is a target of protective immune responses and represents a novel addition to the family of merozoite surface proteins that contain one or more EGF-like domains.

* Corresponding author. Mailing address: Meharry Medical College, Department of Microbiology, 1005 D. B. Todd Blvd., Nashville, TN 37208. Phone: (615) 327-5726. Fax: (615) 327-6072. E-mail: jburns{at}mail.mmc.edu.

Infection and Immunity, November 2000, p. 6189-6195, Vol. 68, No. 11
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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