Williamson J, McLaren D J
J Protozool. 1981 Nov;28(4):460-7. doi: 10.1111/j.1550-7408.1981.tb05320.x.
Phosphatase activity in Trypanosoma rhodesiense has been examined histochemically by light and electron microscopy and by enzymatic assay in homogenate fractions. Using a method with lead as capture ion, acid phosphatase was found in lysosome-like vesicles and in the flagellar pocket. No alkaline adenosine triphosphatase (ATPase) was detectable by this method. Direct assay of p-nitrophenylphosphatase activity in homogenate fractions showed that acid phosphatase activity was strongly membrane-bound, but that activity at pH 9 was minimal in both soluble and particulate fractions. "Endogenous" ATPase activity was localized specifically and reproducibly in the mitochondrial membranes and under the plasma membrane of he flagellum. This nonenzymic reaction product could not be eradicated by glycerol extraction or glucose depletion. Unlike the membrane staining, which was manifest only after lead treatment, heat-resistant electron-dense material was found in the matrix of lysosomal vesicles in trypanosomes fixed in glutaraldehyde only and not subjected to further treatment with heavy metal reagents. X-ray emission analysis showed the presence of calcium and phosphorus, indicating that the matrix might have a phosphate storage function.
通过光学显微镜和电子显微镜以及匀浆组分中的酶活性测定，对罗德西亚锥虫中的磷酸酶活性进行了组织化学检查。使用以铅作为捕获离子的方法，在溶酶体样小泡和鞭毛袋中发现了酸性磷酸酶。用这种方法未检测到碱性腺苷三磷酸酶（ATP酶）。对匀浆组分中对硝基苯磷酸酶活性的直接测定表明，酸性磷酸酶活性与膜紧密结合，但在pH 9时，可溶性和颗粒性组分中的活性都很低。“内源性”ATP酶活性特异性地且可重复地定位于线粒体膜和鞭毛质膜下。这种非酶反应产物不能通过甘油提取或葡萄糖耗尽而消除。与仅在铅处理后才出现的膜染色不同，在仅用戊二醛固定且未用重金属试剂进一步处理的锥虫溶酶体小泡基质中发现了耐热电子致密物质。X射线发射分析表明存在钙和磷，这表明基质可能具有磷酸盐储存功能。
