Localization of acetylated alpha-tubulin in Tritrichomonas foetus and Trichomonas vaginalis.
We used monoclonal antibodies specific for acetylated and nonacetylated alpha-tubulin to detect and to localize microtubules containing acetylated alpha-tubulin (stable microtubules) in the pathogenic protozoa Tritrichomonas foetus and Trichomonas vaginalis. SDS-PAGE analysis showed that tubulin is a major protein of both parasites, being enriched in cytoskeletal preparations of whole cells extracted with Triton X-100. The monoclonal antibodies, which recognize all isoforms of alpha-tubulin (B-5-1-2) and only acetylated alpha-tubulin (6-11B-1), bind to the tubulin of T. foetus and T. vaginalis as seen by immunoblotting. Tubulin-containing structures were localized using immunofluorescence microscopy and transmission electron microscopy of the whole cytoskeleton previously incubated in the presence of the anti-tubulin antibodies and a second antibody-gold complex, and then processed using the negative staining or replica techniques. The results obtained indicate that, in addition to the flagellar microtubules, those which form the peltar-axostyle system represent stable microtubules containing acetylated alpha-tubulin.
Batista
C M
CM
Laboratório de Microscopia Eletrônica e Ultraestrutura Celular, Universidade Federal do Rio de Janeiro, Brasil.
Benchimol
M
M
Cunha e Silva
N L
NL
de Souza
W
W
D000107:Acetylation; D000818:Animals; D000911:Antibodies, Monoclonal; D003078:Colchicine / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007150:Immunohistochemistry; D008854:Microscopy, Electron / Q000379:methods; D008870:Microtubules / Q000187:drug effects / Q000648:ultrastructure; D014246:Trichomonas vaginalis / Q000032:analysis*; D014318:Tritrichomonas / Q000032:analysis*; D014404:Tubulin / Q000032:analysis* / Q000378:metabolism; D014747:Vinblastine / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D014404:Tubulin; D014747:Vinblastine; D003078:Colchicine