70 KDA HEAT SHOCK PROTEINS AND THE HOMOLOGOUS UNCOATING ATPASE

我们实验室的总体重点是70个 kDa热休克蛋白在正常细胞过程和 热休克现象。 尽管我们最终计划 为了研究这些蛋白质在酵母中的功能，我们 通过调查的关系来接近这个问题， 热休克蛋白转化为同源蛋白， 哺乳动物中参与内吞作用的ATP酶 细胞 这种蛋白质从网格蛋白包被的囊泡中剥离网格蛋白， 这个过程需要ATP。 我们的实验室分离出了 囊泡，网格蛋白，网格蛋白衍生的篮，和未涂层 牛脑ATP酶。 我们开发了一种检测方法， 定量测量未被膜的ATP酶 从包被囊泡和网格蛋白篮中除去网格蛋白。 与其他研究人员报道的相反， 未包被的ATP酶催化地从这些细胞中除去网格蛋白。 结构，我们的初步结果表明，它可能会采取行动， 化学计量学上。 我们目前正在调查这是否是 事实上，如果是这样的话，这种化学计量效应如何与 蛋白质的ATP酶活性。

The overall focus of our laboratory is on the function of the 70 kDa heat shock proteins in both normal cellular processes and in the heat shock phenomenon. Although we ultimately plan to investigate the function of these proteins in yeast, we are approaching this question by investigating the relationship of the heat shock proteins to a homologous protein, the uncoating ATPase which appears to be involved in endocytosis in mammalian cells. This protein strips clathrin from clathrin coated vesicles in a process which requires ATP. Our laboratory has isolated coated vesicles, clathrin, clathrin-derived baskets, and the uncoating ATPase from bovine brain. We have developed an assay to quantitatively measure the extent to which the uncoating ATPase removes clathrin from both coated vesicles and clathrin baskets. In contrast to other researchers who have reported that the uncoating ATPase catalytically removes clathrin from these structures, our preliminary results indicate that it may act stoichiometrically. We are currently investigating whether this is indeed the case, and if so, how this stoichiometric effect relates to the ATPase activity of the protein.