Wiser M F, Wood P A, Eaton J W, Sheppard J R
J Cell Biol. 1983 Jul;97(1):196-201. doi: 10.1083/jcb.97.1.196.
Normal and Plasmodium berghei (NYU-2 strain)-infected murine erythrocytes display substantially different patterns of plasma membrane phosphoproteins phosphorylation. Intact erythrocytes (normal and parasite infected) incubated with 32Pi and isolated washed erythrocyte plasma membranes incubated with gamma-32P-ATP were analyzed for phosphoproteins by SDS PAGE and autoradiography. Two new phosphoproteins of molecular weight 45,000 (pp45) and 68,000 (pp68), which are absent in normal erythrocyte membranes, are associated with the membranes of infected erythrocytes subjected to both intact-cell and isolated-membrane phosphorylation conditions. Two-dimensional gel electrophoresis indicates that pp45 and pp68 are of parasite origin. Partial or complete proteolytic digestion reveals that pp45 is phosphorylated at similar amino acid residues both in intact cells and in isolated membranes. The pp45 phosphoprotein can be detected at as low as 3% parasitemia and its phosphorylation is not affected by 10 microM cAMP, 1 mM Ca2+, or 5 mM EGTA. Extraction of isolated washed plasma membranes with 0.5% Triton X-100 or 0.1 M NaOH indicates that pp45 is detergent insoluble and only partially extractable with NaOH, suggesting that pp45 is closely associated with the host erythrocyte plasma membrane.
正常小鼠红细胞和感染伯氏疟原虫（NYU - 2株）的小鼠红细胞表现出截然不同的质膜磷蛋白磷酸化模式。将完整红细胞（正常和感染寄生虫的）与³²Pi一起孵育，并将分离洗涤后的红细胞质膜与γ - ³²P - ATP一起孵育，通过SDS - PAGE和放射自显影分析磷蛋白。正常红细胞膜中不存在的两种分子量分别为45,000（pp45）和68,000（pp68）的新磷蛋白，与在完整细胞和分离膜磷酸化条件下处理的感染红细胞膜相关。二维凝胶电泳表明pp45和pp68来源于寄生虫。部分或完全蛋白水解消化显示，pp45在完整细胞和分离膜中的相似氨基酸残基处被磷酸化。pp45磷蛋白在低至3%的寄生虫血症水平时即可检测到，其磷酸化不受10微摩尔/升环磷酸腺苷、1毫摩尔/升钙离子或5毫摩尔/升乙二醇双四乙酸的影响。用0.5% Triton X - 100或0.1摩尔/升氢氧化钠提取分离洗涤后的质膜表明，pp45不溶于去污剂，仅部分可被氢氧化钠提取，这表明pp45与宿主红细胞质膜紧密相关。
