IMMUNOLOGY & DISEASES OF THE GASTROINTESTINAL TRACT

免疫学

• 批准号: 3225115
• 负责人: JAMES E. MC GUIGAN
• 金额: $ 21.79万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-01-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3225115
• 关键词: Zollinger Ellison syndrome; adrenergic receptor; affinity chromatography; cholecystokinin; collagenase; complement inhibitors; complement pathway; complement receptor; cyclic AMP; dogs; duodenal ulcer; endocrine pharmacology; gastric inhibitory peptide; gastric mucosa; gastrins; gastrointestinal disorder; gastrointestinal hormones; gastrointestinal pharmacology; hemocyanin; histochemistry /cytochemistry; hormone regulation /control mechanism; human tissue; immunochemistry; intestinal mucosa; laboratory rabbit; liver cells; membrane proteins; messenger RNA; peptide hormone biosynthesis; polyvinyls; protein structure function; radioimmunoassay; radiotracer; receptor binding; secretion; somatostatin; transport proteins; vagotomy; vasoactive intestinal peptide

This research proposal is directed to the study of gastrointestinal regulatory peptides with special emphasis on interactions with their receptors as well as characterization of structure and functional characteristics of cell receptor membrane proteins for these regulatory peptides. Peptides complementary to gastrointestinal regulatory peptides will be synthesized by solid phase peptide synthesis based on mRNA complementarity to the codons of human gastrin I (G17), the carboxyl- terminal gastrin tetrapeptide (G4), the carboxyl-terminal octapeptide of cholecystokinin (CCK8), somatostatin-14 (S14) and the carboxyl-terminal tetradecapeptide of gastrin-releasing peptide (GRP14). Ligand binding properties of these complementary peptides will be assessed for the respective gastrointestinal regulatory peptides by incubation of 125 I- gastrointestinal regulatory peptides in wells of polyvinyl chloride plates coated with the respective complementary peptides. Competition studies will be performed to assess specificity of binding of 125 I- regulatory peptides. Antibodies to complementary peptides will be produced by immunization of rabbits with complementary peptides conjugated to keyhole limpet hemocyanin as carrier protein. Potential binding of these antibodies, which will be purified by immunoaffinity chromatography and radiolabelled with 125 I, to cell membrane receptors (on gastric mucosal parietal cells, gastrin cells and somatostatin cells) will be examined. Cells will be prepared from canine fundic mucosa by collagenase dissociation followed by elutriation. Antibodies to complementary peptides, anticipated to bind to binding sites of membrane receptor proteins will be utilized in immunoaffinity studies to purify cell membrane receptors for gastrin, somatostatin, and gastrin-releasing peptide in order to determine the structure of those membrane receptors. Initial studies will be directed to characterization of membrane receptors for gastrin. In the later stages of the proposed studies, using techniques as described for the study of canine fundic mucosal cells, experiments will also be performed to examine structural and functional characteristics of gastrinoma cell membrane receptors for gastrin-releasing peptide and somatostatin.

该研究计划针对胃肠道的研究 调节肽，特别强调与其相互作用 受体以及结构和功能的表征 这些调节的细胞受体膜蛋白的特征 肽。 与胃肠道调节肽互补的肽 将通过基于mRNA的固相肽合成来合成 与人胃泌素 I (G17) 密码子的互补性，即羧基 末端胃泌素四肽 (G4)，羧基末端八肽 胆囊收缩素 (CCK8)、生长抑素-14 (S14) 和羧基末端 胃泌素释放肽（GRP14）的十四肽。 配体结合 这些互补肽的特性将被评估 通过 125 I- 温育分别获得胃肠道调节肽 聚氯乙烯孔中的胃肠道调节肽 板涂有相应的互补肽。 竞赛 将进行研究以评估 125 I- 结合的特异性 调节肽。 互补肽的抗体将是 通过用互补肽免疫兔子产生 与匙孔血蓝蛋白缀合作为载体蛋白。 潜在的 这些抗体的结合，将通过免疫亲和力进行纯化 层析并用 125 I 放射性标记至细胞膜受体 （对胃粘膜壁细胞、胃泌素细胞和生长抑素细胞） 将接受检查。 细胞将从犬胃底粘膜制备 胶原酶解离，然后淘析。 抗体 互补肽，预计与膜的结合位点结合 受体蛋白将用于免疫亲和研究以纯化 胃泌素、生长抑素和胃泌素释放的细胞膜受体 肽以确定这些膜受体的结构。 初步研究将针对膜的表征 胃泌素受体。 在拟议研究的后期阶段， 使用用于研究犬胃底粘膜的技术 细胞，还将进行实验来检查结构和 胃泌素瘤细胞膜受体的功能特征 胃泌素释放肽和生长抑素。