Adrenal Medullary Enkephalin-Containing Peptides

肾上腺髓质脑啡肽含肽

• 批准号: 8719149
• 负责人: Steven Wilson
• 金额: $ 18.28万
• 依托单位: University of South Carolina at Columbia
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-15 至 1991-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8719149&HistoricalAwards=false
• 关键词: Adrenal; Medullary; Enkephalin; Containing; Peptides

Although the amino acid sequence and the identities of many processing products of proenkephalin are known, little information is available on the rates of proenkephalin synthesis and degradation or on the details of proenkephalin processing and storage. The first aim of this project is to determine the rates of proenkephalin synthesis and degradation. Bovine adrenal medullary chromaffin cells will be incubated with radiolabeled amino acids, and their incorporation into enkephalin sequences in the various proenkephalin fragments will be determined following fractionation by high performance liquid chromatography. The second aim is to determine the pathway of proenkephalin processing. Pulse-chase radiolabeling of enkephalin sequences within chromaffin cell proenkephalin will be used to study this pathway. The third aim is to determine the second messenger(s) responsible for regulation of proenkephalin synthesis and processing. The roles of calcium, cyclic AMP, and other agents, including glucocorticiods, will be evaluated. The forth aim is to determine the relationship between proenkephalin and chromaffin vesicle synthesis. Vesicles from radiolabeled chromaffin cells will be subjected to density gradient centrifugation; fractions will be analyzed for newly synthesized enkephalin sequences and other vesicle components. These studies will elucidate the basic mechanisms of neurotransmitter-hormone synthesis and storage.

虽然已知前脑啡肽的氨基酸序列和许多加工产物的特性，但关于前脑啡肽合成和降解的速率或前脑啡肽加工和储存的细节的信息很少。该项目的第一个目的是确定前脑啡肽的合成和降解速率。牛肾上腺髓质染色质细胞将与放射性标记的氨基酸一起孵育，并在高效液相色谱分离后测定其与脑啡肽片段中脑啡肽序列的结合。第二个目的是确定前脑啡肽加工的途径。脑啡肽序列在染色质细胞内的脉冲追踪放射性标记将用于研究这一途径。第三个目的是确定负责调节前脑啡肽合成和加工的第二信使。将评估钙、环AMP和其他药物（包括糖皮质激素）的作用。第四个目的是确定前脑啡肽与嗜铬蛋白囊泡合成的关系。来自放射性标记的嗜铬细胞的囊泡将进行密度梯度离心；将分析新合成的脑啡肽序列和其他囊泡成分。这些研究将阐明神经递质激素合成和储存的基本机制。