Samuelson J C, Caulfield J P
J Cell Biol. 1982 Aug;94(2):363-9. doi: 10.1083/jcb.94.2.363.
Schistosomula of Schistosoma mansoni were labeled by oxidation with galactose oxidase or with periodate followed by reduction with NaB3H4 to study the loss of the surface membrane of these parasites in vitro. Grain counts of light microscope autoradiographs (LMARG) of radiolabeled schistosomula show that both galactose oxidase and periodate specifically label the surface of the organisms. Galactose oxidase labels 11 glycoproteins on the surface of skin and mechanical schistosomula, ranging in apparent molecular weight from 17,000 to greater than 105,000. These glycoproteins are lost from the surface of schistosomula with a halftime of 10-15 h in culture in defined medium. Most of these glycoproteins appear to be shed intact from the surface of the schistosomula rather than endocytosed and degraded, because greater than 50% of each of the lost proteins can be recovered by trichloroacetic acid precipitation of the culture medium and because there is no internalization of the radiolabels into cultured schistosomula examined by LMARG. In addition to glycoproteins, periodate labels at least seven glycolipids on the surface of mechanical schistosomula. After culture for 15 h, more than half of each of these periodate-labeled proteins and lipids are lost from the schistosomula, and their abundance relative to each other remains similar to that of freshly labeled organisms. Since both proteins and lipids are lost from the surface of the schistosomula at the same rate, we believe that we are observing a general loss of the parasite surface membrane.
用半乳糖氧化酶氧化或高碘酸盐氧化后用硼氢化钠还原对曼氏血吸虫童虫进行标记,以研究这些寄生虫体外表面膜的丢失情况。放射性标记童虫的光学显微镜放射自显影片(LMARG)的颗粒计数显示,半乳糖氧化酶和高碘酸盐都能特异性标记虫体表面。半乳糖氧化酶标记皮肤型和机械损伤型童虫表面的11种糖蛋白,表观分子量范围从17,000到大于105,000。在限定培养基中培养时,这些糖蛋白从童虫表面丢失的半衰期为10 - 15小时。这些糖蛋白中的大多数似乎是从童虫表面完整脱落而不是被内吞和降解,这是因为通过对培养基进行三氯乙酸沉淀可回收超过50%的每种丢失蛋白,并且通过LMARG检查发现放射性标记没有内化到培养的童虫中。除了糖蛋白,高碘酸盐还标记机械损伤型童虫表面至少七种糖脂。培养15小时后,这些高碘酸盐标记的蛋白和脂质中的每一种都有超过一半从童虫中丢失,并且它们彼此之间的丰度与新标记虫体的相似。由于蛋白和脂质以相同速率从童虫表面丢失,我们认为我们观察到的是寄生虫表面膜的普遍丢失。