Mukhopadhyay N K, Saha A K, Lovelace J K, Da Silva R, Sacks D L, Glew R H
Department of Microbiology, School of Medicine, University of Pittsburgh, Pennsylvania 15261.
J Protozool. 1988 Nov;35(4):601-7. doi: 10.1111/j.1550-7408.1988.tb04158.x.
Promastigotes from log phase and stationary phase cultures of Leishmania donovani, L. braziliensis panamensis, L. tropica, L. major, and L. mexicana amazonensis were analyzed for their content of protein kinase and acid phosphatase activities. Cell surface, histone-specific protein kinase activity was 1.3- to 2.8-fold higher in stationary phase cells of all species except for L. tropica in which the activities of stationary and log phase cells were equal; L. mexicana amazonensis had the highest histone-specific protein kinase activity and L. donovani the lowest. When viable, motile promastigotes of all five species were incubated for 10 min with [gamma-32P]ATP and Mg2+ (10 mM) in the absence of exogenous histone acceptor; about one dozen proteins were phosphorylated in each case. Both log phase and stationary phase promastigotes of all five species extensively phosphorylated a 50-kDa protein that had the mobility of tubulin. Incubation of pure calf brain tubulin with [gamma-32P]ATP and purified L. donovani protein kinase resulted in extensive phosphorylation of the former. Highly infective metacyclic forms (PNA-) of L. major, isolated from a stationary culture using the peanut agglutinin (PNA), contained eight times more histone-specific protein kinase activity than noninfective log phase cells (PNA+). The PNA- and PNA+ forms of L. major both phosphorylated a 50-kDa protein when incubated with [gamma-32P]ATP and magnesium or manganese ions (10 mM); the 50-kDa protein was precipitated by anti-tubulin rabbit antibodies. Extracts of all five species contained large amounts of acid phosphatase activity.(ABSTRACT TRUNCATED AT 250 WORDS)
对来自杜氏利什曼原虫、巴拿马利什曼原虫、热带利什曼原虫、硕大利什曼原虫和亚马逊利什曼原虫对数期和平稳期培养物的前鞭毛体进行了蛋白激酶和酸性磷酸酶活性含量分析。除热带利什曼原虫外，所有物种的静止期细胞表面组蛋白特异性蛋白激酶活性比对数期细胞高1.3至2.8倍，热带利什曼原虫静止期和对数期细胞的活性相等；亚马逊利什曼原虫的组蛋白特异性蛋白激酶活性最高，杜氏利什曼原虫最低。当所有五个物种的有活力、可运动的前鞭毛体在没有外源组蛋白受体的情况下与[γ-32P]ATP和Mg2+（10 mM）孵育10分钟时；每种情况下约有十二种蛋白质被磷酸化。所有五个物种的对数期和平稳期前鞭毛体都广泛磷酸化了一种具有微管蛋白迁移率的50 kDa蛋白质。将纯小牛脑微管蛋白与[γ-32P]ATP和纯化的杜氏利什曼原虫蛋白激酶一起孵育，导致前者大量磷酸化。从使用花生凝集素（PNA）的静止培养物中分离出的硕大利什曼原虫的高感染性循环后期形式（PNA-），其组蛋白特异性蛋白激酶活性比非感染性对数期细胞（PNA+）高八倍。当与[γ-32P]ATP和镁或锰离子（10 mM）一起孵育时，硕大利什曼原虫的PNA-和PNA+形式都磷酸化了一种50 kDa蛋白质；50 kDa蛋白质被抗微管蛋白兔抗体沉淀。所有五个物种的提取物都含有大量的酸性磷酸酶活性。（摘要截断于250字）
