MN2+ -PHOSPHOLIPID STIM PK AND LEUKEMIC DIFFERENTIATION

该项目涉及一种新的生物化学和细胞生理学 识别的蛋白激酶，称为“PK-P”，用于 磷脂调节的蛋白激酶及其主要内源性底物 第73页。这个系统在小鼠和人类白血病细胞中都可以检测到， 其中它可能在多克隆刺激因子(IL-3)信号中发挥作用 转导和化学诱导分化。第73页一直是 纯净的，并将进一步表征，关于 观察到的蛋白水解性；它的相互作用 以磷脂和二价阳离子为基础，以蛋白激酶为基础 底物；以及通过部分氨基酸测序确定的结构 和免疫学分析。PK-P也被发现很容易 磷酸化核糖体蛋白S6。这一反应将进一步 量化并与其他激酶的磷酸化进行比较，及其 确定了对体外翻译的影响。前体、其他亚基 或相互作用的活动，或调节PK-P的修饰 已确认身份。PK-P/pp73体系对化学物质的响应模式 和HL-60人骨髓细胞的生物分化诱导剂 白血病细胞株的建立及IL-3对DA-1小鼠白血病细胞的作用 LINE将使用直接测量蛋白质和 蛋白质的磷酸化水平和亚细胞分布 免疫印迹技术。这些组分与膜的相互作用 也将对这些细胞进行各种这样的处理进行研究。 分子生物学研究的目的是克隆人的cDNA3基因。 将进行PK-P，以便PK-P的完整一级结构可以 最终会被决定。

This project deals with the biochemistry and cellular physiology of a newly identified protein kinase, referred to as "PK-P", for phospholipid-modulated protein kinase, and its major endogenous substrate pp73. This system is detectable in murine and human leukemic cells, wherein it may play a role in multi-colony stimulating factor (IL-3) signal transduction and chemically inducible differentiation. pp73 has been purified, and will be characterized further, with respect to the proteolytic activity which it has been observed to exhibit; its interaction with phospholipids and divalent cations, and with protein kinases as a substrate; and its structure as determined by partial amino acid sequencing and immunologic analysis. PK-P has also been found to readily phosphorylate ribosomal protein S6. This reaction will be further quantitated and compared to phosphorylation by other kinases, and its effects upon in vitro translation determined. Precursors, other subunits or interacting activities, or modifications modulating PK-P will be identified. The pattern of response of the PK-P/pp73 system to chemical and biologic differentiation inducing agents of the HL-60 human myeloid leukemic cell line, and to IL-3 treatment of the DA-1 murine leukemic cell line will be further characterized using direct measurement of protein and phosphorylation levels and subcellular distribution by protein immunoblotting techniques. Interaction of these components with membranes from such cells subjected to various such treatments will also be studied. Molecular biologic studies aimed towards the cloning of the cDNA gene for PK-P will be undertaken, so that the complete primary structure of PK-P can ultimately be determined.