Schwartzman J D, Krug E C
J Protozool. 1986 Nov;33(4):576-8. doi: 10.1111/j.1550-7408.1986.tb05665.x.
Monoclonal antibodies specific for mammalian beta-tubulin recognized the microtubule cytoskeleton of the flagellated protozoon Trichomonas vaginalis. Of seven antibodies, two demonstrated the axostyle, costa, recurrent flagellum, and anterior flagella by indirect immunofluorescence microscopy. The remaining five stained a hazy reticular pattern in the cytoplasm of formaldehydefixed, detergent-extracted organisms. Western immunoblots of whole T. vaginalis extracts treated with protease inhibitors and electrophoresed on polyacrylamide gels containing sodium dodecyl sulfate showed a major band at molecular weight 50,000 when probed with only one of the antibodies which stained the axial cytoskeleton. The antibodies which stained only the cytoplasm showed a different western blot pattern with a major doublet band at MW 58,000-60,000. Another antibody, which stained both the axial cytoskeleton and the reticular cytoplasmic pattern showed major bands at MW 58,000-60,000 and also at MW 40,000-42,000. The recognition of microtubule populations in T. vaginalis by these monoclonal antibodies was different than we found earlier with Leishmania donovani and Toxoplasma gondii, where all seven antibodies recognize cytoskeletal microtubules and produce western blots characteristic of tubulin. Only one of these seven antibodies recognizes tubulin in T. vaginalis by immunoblot. The microtubules of T. vaginalis do not demonstrate all epitopes recognized by monoclonal antibodies specific for mammalian beta-tubulin; one of the antibodies appears to recognize an epitope which is morphologically associated with microtubules but does not have the characteristic MW of tubulin.
针对哺乳动物β-微管蛋白的单克隆抗体识别了有鞭毛的原生动物阴道毛滴虫的微管细胞骨架。在七种抗体中，两种通过间接免疫荧光显微镜显示出轴柱、肋、回鞭毛和前鞭毛。其余五种在经甲醛固定、去污剂处理的生物体细胞质中染出模糊的网状图案。用蛋白酶抑制剂处理并在含十二烷基硫酸钠的聚丙烯酰胺凝胶上电泳的阴道毛滴虫全提取物的蛋白质免疫印迹显示，仅用一种染轴细胞骨架的抗体检测时，在分子量50,000处有一条主要条带。仅染细胞质的抗体显示出不同的蛋白质免疫印迹模式，在分子量58,000 - 60,000处有一条主要的双峰条带。另一种既染轴细胞骨架又染细胞质网状图案的抗体在分子量58,000 - 60,000以及分子量40,000 - 42,000处显示出主要条带。这些单克隆抗体对阴道毛滴虫微管群体的识别与我们早期在杜氏利什曼原虫和刚地弓形虫中发现的不同，在那里所有七种抗体都识别细胞骨架微管并产生微管蛋白特征性的蛋白质免疫印迹。这七种抗体中只有一种通过免疫印迹识别阴道毛滴虫中的微管蛋白。阴道毛滴虫的微管并未显示出针对哺乳动物β-微管蛋白的单克隆抗体所识别的所有表位；其中一种抗体似乎识别一种在形态上与微管相关但不具有微管蛋白特征分子量的表位。
