ROLES OF PHOSPHOLIPID AND CA2+ IN BRAIN

此更新应用程序的总体目标主要包括 的分子、免疫学、药理学和调节方面， 最近鉴定的磷脂敏感性钙依赖性蛋白激酶 （PL-Ca-PK）系统。 具体目标如下：（1） PL-Ca-PK酶 将尝试改进化合物的纯化。 猪或牛脑中的酶。 同样，单克隆抗体的生产 抗体的酶将得到改善，和分布的 将用免疫细胞化学方法研究脑中的酶。 (2)内源 印刷受体. 髓鞘碱性蛋白（MBP）的磷酸化位点， 将测定PL-Ca-PK。 一旦完成，最小的结构 将探讨PL-Ca-PK底物特异性的决定因素 使用合成寡肽。 磷酸化MBP和 其片段或肽对它们诱导 将探索实验性过敏性脑脊髓炎。 除了 MBP，脑中PL-Ca-PK的许多其他内源性底物， 进一步调查。 (3)PL-Ca-PK系统的调节。 法规 各种试剂的酶活性，磷酸化状态， 内源性底物与脑内PL-Ca-PK可能的“易位” 将被调查。 (4)一种新的S-100调节的蛋白磷酸化 系统 S-100调节的酶（蛋白激酶X）及其19 K Mr 将进一步纯化和表征大鼠脑中底物蛋白。 希望该提案将产生新的知识， 磷脂、钙离子和S-100对脑生物学和病理学的调节 蛋白质磷酸化反应。

The overall goals of this renewal application cover primarily the molecular, immunological, pharmacological and regulatory aspects of the recently-identified phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK) system in brain. The specific aims are as follows: (1) PL-Ca-PK, the enzyme. Attempts will be made to improve the purification of the enzyme from pig or bovine brain. Similarly, production of monoclonal antibodies to the enzyme will be improved, and the distribution of the enzyme in brain will be studied immunocytochemically. (2) Endogenous substrates. The sites of phosphorylation in myelin basic protein (MBP) by PL-Ca-PK will be determined. Once this is done, the minimal structural determinants of the substrate specificity for PL-Ca-PK will be explored using synthetic oligopeptides. The effects of phosphorylation of MBP and its fragments or peptides on their ability or inability to induce experimental allergic encephalomyelitis will be explored. In addition to MBP, a number of other endogenous substrates for PL-Ca-PK in brain will be further investigated. (3) Regulation of PL-Ca-PK system. Regulations by various agents of the enzyme activity, the phosphorylation state of endogenous substrates and possible "translocation" of PL-Ca-PK in brain will be investigated. (4) A new S-100-modulated protein phosphorylation system. The S-100-modulated enzyme (protein kinase X) and its l9K Mr substrate protein in rat brain will be further purified and characterized. It is hoped that the proposal will produce new knowledge regarding regulation of brain biology and pathology by phospholipid, Ca2+ and S-100 protein via protein phosphorylation reaction.