SECRETORY SALIVARY PROTEIN SPECIFIC PROTEIN KINASES

分泌性唾液蛋白特异性蛋白激酶

• 批准号: 2132241
• 负责人: MARK S LAMKIN
• 金额: $ 10.23万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-04-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2132241
• 关键词: Macaca fascicularis; SDS polyacrylamide gel electrophoresis; affinity chromatography; antibody; chemical kinetics; complementary DNA; enzyme activity; enzyme structure; enzyme substrate; genetic library; histidine; human subject; molecular cloning; molecular site; oligonucleotides; parotid gland; peptides; proline; protein kinase; protein purification; protein sequence; salivary glands; secretory protein; tyrosine

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.

本研究的总体目标是表征蛋白质 存在于大唾液腺中的激酶，其磷酸化分泌 唾液蛋白 唾液基质的磷酸化形式， 富含脯氨酸的酸性蛋白质、statherin和histatin 1，特别是， 在维持获得的釉质表膜中是重要的， 牙齿完整性 将用各种肽测定激酶活性 底物中磷酸丝氨酸周围的序列， 富含脯氨酸的酸性蛋白质，statherin和histatin 1。 这些 活性将从食蟹猴腮腺中纯化 通过连续色谱技术。 底物特异性， 核苷酸需求、金属需求和pH依赖性将是 对于每种纯化的激酶测定。 此外，氨基末端 将确定每种激酶的序列。 这些序列将用于 开发寡核苷酸探针，除了抗体， 用于筛选猕猴腮腺cDNA表达文库。 完整的一级结构将由渐进式 寡核苷酸测序。 这些序列将与 现有的序列数据库。 此外，两人的存在 已知丝氨酸/苏氨酸激酶的共有序列将被 测定 来自氨基酸和 寡核苷酸测序研究将用于分析蛋白质 已建立的数据库中的序列。 这些研究对于理解法规和 的合成、磷酸化和分泌的协调， 不可逆磷酸化的蛋白质，如唾液 磷蛋白、维生素E和磷蛋白。