VIP, VIP RECEPTORS AND THE INTESTINAL EPITHELIAL CELL

VIP、VIP 受体和肠上皮细胞

• 批准号: 3463556
• 负责人: TOAN D. NGUYEN
• 金额: $ 7.45万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3463556
• 关键词: Golgi apparatus; affinity chromatography; antibody; endoplasmic reticulum; gastrointestinal epithelium; gel filtration chromatography; glycosylation; laboratory mouse; laboratory rabbit; laboratory rat; liposomes; membrane lipids; monoclonal antibody; neoplastic cell culture for noncancer research; protein biosynthesis; receptor; tissue /cell culture; vasoactive intestinal peptide

The effects of vasoactive intestinal peptide (VIP) on intestinal secretion have been well characterized. Clinically, tumors producing VIP are responsible for the syndrome of watery diarrhea, hypokalemia and gastric achlorhydria described by Verner and Morrison. However, the interaction between VIP and its receptor on the target cell, the first step responsible for specific signal transduction, has only recently been examined. With the groundwork laid by our previous studies on the structural on the intestinal VIP receptor, we plan to: 1. Raise monoclonal and polyclonal antibodies against the VIP receptor using the purest receptor preparation available (see below). 2. Purify the VIP receptor relying mainly on gel filtration and affinity chromatography (using either VIP or antibodies against the receptor). If required, reconstitute the purified receptor into liposomes to define its function. 3. Using different endoglycosidases, study the degree and type of glycosylation of the VIP receptor located on the ER-Golgi and laterobasal membranes of the enterocyte. With the panel of monoclonal antibodies described in 1, attempt to correlate a function or structure to each domain of the receptor. 4. Study the synthesis, glycosylation, vectorial processing and transport of the metabolically radiolabeled VIP receptor in pulse chase experiments using the isolated intestinal loop. 5. With the intestinal loop or the human colonic adenocardinoma cell line, HT 29, study the cellular cycle of the receptor (internalization, degradation, recycling).

血管活性肠肽（VIP）对肠道的影响 分泌的特征很好。 临床上是肿瘤 生产VIP负责水状综合征 腹泻，低钾血症和胃ach术 Verner和Morrison。 但是，VIP和 它的受体在目标细胞上，第一步负责 特定的信号转导，直到最近才被检查。 我们先前关于结构的研究奠定了基础 在肠道受体上，我们计划： 1。提出针对VIP的单克隆和多克隆抗体 使用最纯的受体制剂的受体（请参阅 以下）。 2。纯化的VIP受体主要依赖于凝胶过滤和 亲和色谱法（使用VIP或抗体 受体）。 如果需要，将纯化的受体重新构成 脂质体定义其功能。 3。使用不同的内糖苷酶，研究的程度和类型 位于ER-Golgi上的VIP受体的糖基化和 肠肠细胞的后叶膜。 带有面板 1中描述的单克隆抗体试图将A相关 受体每个域的功能或结构。 4。研究合成，糖基化，媒介物加工和 脉搏中代谢放射性标记的VIP受体的转运 使用分离的肠环的追逐实验。 5。带有肠环或人类结肠腺癌 细胞系，HT 29，研究受体的细胞周期 （内部化，退化，回收）。