Characterization of cDNAs encoding adhesin proteins involved in trichomonas vaginalis cytoadherence
Arroyo, R.; Engbring, J.; Nguyen, J.; Musatovova, O.; López, O.; Lauriano, C.; Alderete, J.F.
Archives of Medical Research 26(4): 361-369
1995
ISSN/ISBN: 0188-4409 PMID: 8555730 Document Number: 450577
A cDNA library was constructed from mRNA of Trichomonas vaginalis grown in a high-iron medium. 3 cDNA clones for adhesin AP65, one for AP51 and 6 for AP33 were recognized with polyclonal antiserum or monoclonal antibody raised against these adhesins. No cross-hybridization among cDNA of the 3 adhesions was observed in Southern analysis, confirming restriction mapping analysis of representative cDNAs. Southern analysis probed with representative cDNAs of the adhesins indicated multiple copies for each of the 3 adhesin genes. Northern analysis showed transcripts of 1.8 kb, 1.4 kb and 0.9 kb for AP65, AP51 and AP33, respectively. Consistent with adhesin expression, mRNAs for these adhesins were detected only when parasites were grown in high-iron conditions. Specific antibodies eluted from E. coli expressing recombinant proteins reacted only with the respective parasite adhesins. Recombinant proteins bound to fixed HeLa cells and competed with radiolabelled trichomonad adhesins for binding. Although proteinase activity is required for cytoadherence, recombinant proteins showed no detectable proteinase activity. Data showed that recombinant proteins from these clones exhibited characteristics of the trichomonad adhesins.