RENAL KALLIKREIN-BINDING PROTEIN IN HYPERTENSION

这项建议的目的是研究肾脏的功能作用。 (组织)高血压中的激肽释放酶结合蛋白。组织激肽释放酶是 一组同源丝氨酸蛋白酶，由一个多基因家族和 参与生物活性多肽和激素的加工。激肽释放酶 其产品在细胞内稳态和血液中起着重要作用 压力调节。虽然生物化学和调控 激肽释放酶基因在正常和疾病状态下的表达 经过广泛研究的，关于调节因素的信息 激肽释放酶的作用非常有限。我们最近确定了一部小说 大鼠和人形成的不可逆的激肽释放酶结合蛋白 组织激肽释放酶复合体。大鼠和人的结合蛋白 与组织激肽释放酶形成不可逆的复合体。结合蛋白 在自发性高血压大鼠(SHR)中存在缺陷，这表明它 可能是SHR高血压表型的一个促成因素。 本研究的具体目的是：1)提纯和表征 激肽释放酶结合蛋白；研制单抗和多克隆抗体 为了确定激肽释放酶的分布、数量和定位- 通过结合分析、放射免疫分析、Western blotting、 免疫组织化学和生理学研究；2)阐明 激肽释放酶结合蛋白在激肽释放酶调节中的作用 体内清除和分解代谢研究的生物利用度；3)分离 大鼠肝脏激肽释放酶结合蛋白基因的克隆 文库，并使用这些克隆来鉴定和定位激活素结合 Northern、斑点印迹和原位cDNAm RNA检测蛋白基因表达 4)分离和鉴定编码该基因的基因。 从大鼠基因组文库中提取激肽释放酶结合蛋白，并鉴定 参与组织特异性的潜在顺式和反式作用元件 用缺失分析和核蛋白结合方法表达该基因 检测；5)分析相关的限制性片段长度多态性 携带激肽释放酶结合蛋白基因的遗传性高血压动物 模特们。从这些研究中获得的信息应该会提供更好的 激肽释放酶-激肽系统在高血压中的作用。

The objective of this proposal is to study the functional role of renal (tissue) kallikrein-binding protein in hypertension. Tissue kallikrein are a group of homologous serine proteinases, encoded by a multigene family and involved in the processing of bioactive peptides and hormones. Kallikrein and its products play important roles in cellular homeostasis and blood pressure regulation. Although the biochemistry and regulation of kallikrein gene expression in normal and disease states have been extensively studied, information regarding factors which modulate kallikrein function is very limited. We have recently identified a novel kallikreinbinding protein in rat and human which forms and irreversible complex with tissue kallikrein. The binding protein in rat and human which form an irreversible complex with tissue kallikrein. The binding protein is defective in spontaneously hypertensive rats (SHR), suggesting that it may be a contributing factor in the hypertensive phenotype of SHR. The specific aims of this study are: 1) to purify and characterize kallikrein-binding protein; develop monoclonal and polyclonal antibodies for determining distribution, quantities and localization of kallikrein- binding protein by binding assays, radioimmunoassays, Western blotting, immunohistochemistry and for physiological studies; 2) to elucidate the role of kallikrein-binding protein in the regulation of kallikrein bioavailability by in vivo clearance and catabolism studies; 3) to isolate cDna clones encoding the kallikrein-binding protein from a rat liver cDNA library and to use these clones to identify and localize kallikreinbinding protein gene expression by Northern, dot blot and in situ cDNA-mRNA hybridization; 4) to isolate and characterize the gene encoding the kallikreinbinding protein from a rat genomic library and to identify the potential cis- and trans-acting elements involved in tissue-specific expression of the gene by deletion analysis and nuclear protein binding assays; 5) to analyze restriction fragment length polymorphisms associated with the kallikrein-binding protein gene in genetically hypertensive animal models. The information gained form these studies should provide a better understanding of the kallikrein-kinin system's role in hypertension.