The phenotypically variable surface protein of Trichomonas vaginalis has a single, tandemly repeated immunodominant epitope.
Trichomonas vaginalis is a sexually transmitted protozoan parasite that undergoes phenotypic variation for numerous surface proteins. A monoclonal antibody (MAb) was used to isolate an approximately 400-bp cDNA encoding a fragment of an important phenotypically varying immunogen of T. vaginalis (Mr = 270 kDa; P270). The MAb completely inhibited the binding of P270 by antibody in sera of patients and by antibody in monospecific antiserum obtained toward purified P270, indicating that P270 contained only one immunodominant epitope. Hydrophilicity plot analysis of the deduced amino acid sequence of the recombinant protein predicted the hexapeptide sequence DREGRD as the antigenic determinant of P270. Synthetic peptides synthesized to this region demonstrated that the amino acid sequence DREGRD is important for antibody binding. Seven adjacent amino acids also contributed substantially to maximal recognition of the epitope by the MAb. A single transcript of approximately 9.5 kb, a size compatible with the reported Mr of the immunogen, hybridized to the cDNA in Northern blots of total RNA from T. vaginalis. DNA sequence and Southern blot analysis determined the epitope to be encoded by a 339-bp unit, which was found to be tandemly repeated at least 12 times within the single-copy gene. This 12-mer unit would only constitute approximately 50% of the protein, yet it is responsible for all of the serum antibody to the immunogen produced by animals and humans. The epitope sequence was found in all fresh and long-term-grown organisms examined to date, demonstrating the stability and conservation of this gene.
Dailey
D C
DC
Department of Microbiology, University of Texas Health Science Center, San Antonio 78284-7758.
Alderete
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015152:Blotting, Northern; D004247:DNA / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D005798:Genes, Bacterial / Q000276:immunology; D014644:Genetic Variation / Q000235:genetics*; D016056:Immunodominant Epitopes / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D010641:Phenotype; D012313:RNA / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics* / Q000276:immunology; D014246:Trichomonas vaginalis / Q000235:genetics* / Q000276:immunology
D000953:Antigens, Protozoan; D016056:Immunodominant Epitopes; D011994:Recombinant Proteins; D012313:RNA; D004247:DNA