SECRETORY SALIVARY PROTEIN SPECIFIC PROTEIN KINASES

这项研究的总体目标是确定蛋白质的特征。 磷酸化分泌的主要唾液腺中存在激酶 唾液蛋白质。唾液底物的磷酸化形式， 富含脯氨酸的酸性蛋白，特别是组氨酸和组蛋白1， 在维持后天获得的釉质膜和 牙齿的完整性。将用不同的多肽来检测激酶的活性 底物基于磷酸丝氨酸在 富含脯氨酸的酸性蛋白质、他汀类蛋白和组蛋白1。这些 将从猕猴腮腺中提纯活性 通过顺序层析技术。底物专一性， 核苷酸需求、金属需求和pH依赖将是 为每个纯化的酶测定。此外，氨基末端 每一种酶的序列将被确定。将使用这些序列 开发寡核苷酸探针，除了抗体外，还将 用于从猕猴腮腺中筛选出一个表达文库。 完整的初级结构将由递进决定 寡核苷酸测序。这些序列将与 现有的序列数据库。此外，两者的存在 已知的丝氨酸/苏氨酸激酶的通用序列将是 下定决心。来自氨基酸和氨基酸的序列信息 寡核苷酸测序研究将用于蛋白质的分析 已建立的数据库中的序列。 这些研究将是理解该法规和 合成、磷酸化和分泌的协调 不可逆转的磷酸化蛋白质，如唾液 磷蛋白、磷维生素和磷酸角蛋白。

The overall objective of this study is to characterize the protein kinases present in the major salivary glands which phosphorylate secreted salivary proteins. The phosphorylated forms of the salivary substrates, acidic proline-rich proteins, statherin, and histatin 1, in particular, are important in the maintenance of the acquired enamel pellicle and tooth integrity. Kinase activity will be assayed with various peptide substrates based upon the sequences surrounding the phosphoserines in the acidic proline-rich proteins, statherin, and histatin 1. These activities will be purified from the parotid gland of Macaca fascicularis by sequential chromatographic techniques. The substrate specificity, nucleotide requirement, metal requirement, and pH dependence will be determined for each purified kinase. In addition, the amino-terminal sequence of each kinase will be determined. These sequences will be used to develop oligonucleotide probes, which in addition to antibodies, will be used to screen a cDNA expression library from a macaque parotid gland. The complete primary structure will be determined by progressive oligonucleotide sequencing. The sequences will be compared to those in existing sequence databases. In addition, the existence of the two consensus sequences for known serine/threonine kinases will be determined. Sequence information from both amino acid and oligonucleotide sequencing studies will be used for analysis of protein sequences in established databases. Such studies will be necessary for understanding the regulation and coordination of the synthesis, phosphorylation, and secretion of irreversibly phosphorylated proteins such as the salivary phosphoproteins, phosvitins, and phosphophoryns.