MEMBRANE FUNCTION IN PAROTID SECRETORY MECHANISMS

描述：(逐字摘自应用程序)建议的长期目标 研究旨在了解唾液中介导胞吐作用的机制 腺泡细胞。胞吐作用控制着大分子向口腔的输出。 空洞，因此代表了一种提供蛋白质和 维持口腔生理和宿主防御所必需的多肽 并用于启动消化。调查的重点是一家 高度保守和广泛分布的蛋白质称为SCAMPs(对于 分泌载体膜蛋白)，这是由校长发现的 调查员。SAMP驻留在分泌颗粒中及其相关回收 经历胞吐作用的膜载体，以及至少四种不同的SCAMP 已在腺泡细胞中发现，具有不同的分布。功能性 研究得出这样的假设：SCAMP是分子的一部分 胞吐作用所需的机制及其高度保守的结构域 作用是在膜上组织富含磷脂酰肌醇的微域。五个具体 已经制定了一些目标来解决这一假设，最初的重点是 颗粒相关的SCAMP2。首先，高度保守的结构域的突变 全长的SCAMP将被检查为显性抑制作用 在受调控的分泌细胞系中使用条件表达的胞吐作用。 第二，涉及相似条件表达式的补充性研究将 测试SCAMP和反义基因的调控片段是否具有相似的结构 干扰胞吐作用。第三，对重组SCAMP进行纯化， 在磷脂微囊中重组并用光谱技术进行测试 用于与磷脂酰肌醇的相互作用和调节脂质微域。 第四，将使用一系列措施寻找SCAMP的绑定合作伙伴 检测和分析蛋白质相互作用的程序。第五, 将分析小鼠腮腺唾液腺的胞吐作用。 SCAMP1基因已经被消融(从另一个实验室获得)，并在小鼠身上 缺乏SCAMP2，这是一种将被协作进行的基因敲除。已被占用 总而言之，这些研究源于对NIDCR计划的响应 声明，反映了推进基本知识的持续承诺的一部分 了解分子细胞生物学和口腔生理学之间的关系。

DESCRIPTION: (verbatim from application) The long term goal of the proposed research is to understand the mechanisms mediating exocytosis in salivary acinar cells. Exocytosis controls the export of macromolecules to the oral cavity and thus represents a basic mechanism for supplying proteins and peptides that are essential for maintaining oral physiology and host defense and for initiating digestion. The focus of investigations is on a family of highly conserved and broadly distributed proteins known as SCAMPs (for Secretory Carrier Membrane Proteins), which were discovered by the Principal Investigator. SCAMPs reside in secretory granules and related recycling membrane carriers that undergo exocytosis, and at least four different SCAMPs have been identified in acinar cells with distinct distributions. Functional studies have led to the hypothesis that SCAMPs are part of the molecular machinery required for exocytosis and that their most highly conserved domain acts to organize phosphoinositide-rich microdomains in membranes. Five specific aims have been developed to address this hypothesis, initially focusing on the granule-associated SCAMP2. First, mutations in the highly conserved domain of full-length SCAMP will be examined for dominant inhibitory effects on exocytosis using conditional expression in a regulated secretory cell line. Second, complementary studies involving analogous conditional expression will test whether regulatory segments of SCAMP and antisense constructs similarly interfere with exocytosis. Third, recombinant SCAMP will be purified, reconstituted in phospholipid vesicles and tested by spectroscopic techniques for interaction with phosphoinositides and regulation of lipid microdomains. Fourth, the binding partners of SCAMP will be sought using a battery of procedures to detect and analyze protein-protein interactions. Fifth, exocytosis will be analyzed in parotid salivary glands of mice in which the SCAMP1 gene has been ablated (obtained from another laboratory) and in mice lacking SCAMP2, a gene knockout that will be made collaboratively. Taken together, these studies, which originated in response to an NIDCR program announcement, reflect part of an ongoing commitment to advance basic understanding at the interface of molecular cell biology and oral physiology.