Johnson J G, Epstein N, Shiroishi T, Miller L H
J Protozool. 1981 May;28(2):160-4. doi: 10.1111/j.1550-7408.1981.tb02825.x.
Viable merozoites of Plasmodium knowlesi were isolated and the proteins that were labeled on intact merozoites by lactoperoxidase-catalyzed radioiodination were identified. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography of Triton soluble extracts of labeled merozoites demonstrated eight major bands ranging in apparent molecular weight from 150,000 D to 22,000 D. Exposure of intact merozoites to trypsin (10 microgram/ml) for 10 min resulted in the loss of the two highest molecular weight proteins (150,000 D and 105,000 D) and the appearance of two new bands at 70,000 D and 62,000 D. Trypsin treatment under these conditions also removed the receptor(s) for merozoite attachment to erythrocytes. Therefore, these high molecular weight proteins are candidates for the merozoite component that attaches to erythrocytes. There was no evidence that the labeled membrane components were serum or erythrocyte membrane components, two potential contaminants in the preparation. Anti-rhesus erythrocyte antibody did not precipitate labeled merozoite proteins. Furthermore, the immunoprecipitation of labeled merozoite proteins by rhesus anti-merozoite serum was not inhibited by erythrocyte ghosts.
诺氏疟原虫的活裂殖子被分离出来,并且鉴定了通过乳过氧化物酶催化的放射性碘化标记在完整裂殖子上的蛋白质。对标记裂殖子的Triton可溶性提取物进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳和放射自显影,显示出八条主要条带,表观分子量范围从150,000 D到22,000 D。将完整裂殖子暴露于胰蛋白酶(10微克/毫升)10分钟,导致两条分子量最高的蛋白质(150,000 D和105,000 D)消失,并在70,000 D和62,000 D处出现两条新条带。在这些条件下的胰蛋白酶处理也去除了裂殖子附着于红细胞的受体。因此,这些高分子量蛋白质是裂殖子附着于红细胞的成分的候选者。没有证据表明标记的膜成分是血清或红细胞膜成分,这是制备过程中的两种潜在污染物。抗恒河猴红细胞抗体不会沉淀标记的裂殖子蛋白质。此外,恒河猴抗裂殖子血清对标记裂殖子蛋白质的免疫沉淀不受红细胞血影的抑制。