Ruben L, Haghighat N, Campbell A
Department of Biological Sciences, Southern Methodist University, Dallas, Texas 75275.
Exp Parasitol. 1990 Feb;70(2):144-53. doi: 10.1016/0014-4894(90)90095-t.
The present study was undertaken to evaluate changes in the complement of calmodulin-binding proteins which accompany cyclical differentiation in Trypanosoma brucei. An [125I]trypanosome calmodulin overlay procedure was used to detect calmodulin-binding proteins with Mr of 126,000 and 106,000 that were present in homogenates of slender bloodstream froms but were absent in procyclic culture forms. Competition assays with unlabeled bovine brain or trypanosome calmodulins indicated that the developmentally regulated proteins associated with calmodulins from either source. Moreover, [125I]bovine brain calmodulin associated with the same proteins as trypanosome calmodulin. Homogenates of T. evansi exhibited the same pattern of calmodulin-binding activity as T. brucei slender bloodstream forms; however, T. cruzi and Leishmania tarentolae contained distinct patterns of calmodulin-binding activity. Mouse serum contained no detectable binding proteins while mouse brain contained predominantly proteins of Mr 210,000, 60,000, and 49,000 which were associated with the trypanosome calmodulin probe. The developmentally regulated calmodulin-binding proteins from T. brucei were in the 10,000g pellet. We conclude that the cellular complement of calmodulin-binding proteins varies during the trypanosome life cycle.
本研究旨在评估布氏锥虫周期性分化过程中钙调蛋白结合蛋白的变化。采用[125I]锥虫钙调蛋白覆盖法检测到,在细长型血流型虫体匀浆中存在分子量为126,000和106,000的钙调蛋白结合蛋白,而在原循环培养型中则不存在。用未标记的牛脑或锥虫钙调蛋白进行竞争分析表明,发育调控蛋白与来自任一来源的钙调蛋白相关。此外,[125I]牛脑钙调蛋白与锥虫钙调蛋白结合的是相同的蛋白。伊氏锥虫的匀浆表现出与布氏锥虫细长型血流型相同的钙调蛋白结合活性模式;然而,克氏锥虫和热带利什曼原虫含有不同的钙调蛋白结合活性模式。小鼠血清中未检测到结合蛋白,而小鼠脑中主要含有分子量为210,000、60,000和49,000的蛋白,这些蛋白与锥虫钙调蛋白探针相关。布氏锥虫发育调控的钙调蛋白结合蛋白存在于10,000g的沉淀中。我们得出结论,在锥虫生命周期中,钙调蛋白结合蛋白的细胞组成会发生变化。