THE CART RECEPTOR

购物车接收器

• 批准号: 8357488
• 负责人: MICHAEL J KUHAR
• 金额: $ 2.06万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357488
• 关键词: Affinity; Binding; Binding Sites; Cell membrane; Cycloheximide; Dactinomycin; Funding; G-Protein-Coupled Receptors; GTP gamma S; Gene Expression Regulation; Grant; Hormones; Ligands; Link; National Center for Research Resources; Neurotransmitters; PC12 Cells; Peptides; Pertussis Toxin; Physiological; Primates; Principal Investigator; Protein Biosynthesis; RNA chemical synthesis; Research; Research Infrastructure; Resources; Source; Structure; United States National Institutes of Health; cocaine- and amphetamine-regulated transcript protein; cost; inhibitor/antagonist; pituitary adenylate cyclase activating polypeptide; postsynaptic; receptor; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. CART peptides are peptide neurotransmitters and hormones that are involved in many different physiological responses. While much is known about the peptides regarding their structure, processing and gene regulation, less is known about their postsynaptic actions and receptors. Using 125I-CART 61-102 as a ligand and unlabeled CART 61-102 or CART 55-102 as displacers, high-affinity specific binding was detected in PC12 cells. Differentiation of the PC12 cells increased specific binding several-fold. The increase in specific binding found after differentiation was inhibited by actinomycin D and cycloheximide, suggesting that the increase in specific binding was dependent on RNA and protein synthesis. CART 1-27, a peptide that has never been shown to elicit responses, did not displace 125I-CART61-102 binding, nor did more than 20 other peptides that were examined. Surprisingly, however, PACAP 1-38 and PACAP 6-38 were found to be low-affinity inhibitors of CART binding. CART treatment increased binding of 35S [gamma]GTP-S to PC12 cell membranes. Moreover, CART treatment of intact PC12 cells elicited robust increases in phospho-ERK in a manner that was increased with differentiation, blocked by pertussis toxin and antagonized by PACAP 6-38. These findings suggest that the CART binding site in PC12 cells reflects a G protein-coupled receptor linked with Gi/o, and also demonstrate that PACAP 6-38 may be useful as a CART receptor antagonist.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 推车肽是参与许多不同生理反应的肽神经递质和激素。尽管对肽的结构，加工和基因调节知之甚少，但对它们的突触后作用和受体的了解却少。 将125i-Cart 61-102用作配体和未标记的CART 61-102或CART 55-102作为位移，在PC12细胞中检测到高亲和力特异性结合。 PC12细胞的分化增加了特异性结合几倍。 放线霉素D和环己酰亚胺抑制了分化后发现的特异性结合的增加，这表明特异性结合的增加取决于RNA和蛋白质的合成。 CART 1-27是一种从未显示出引起反应的肽，并未取代125i-Cart61-102结合，也没有进行检查的其他20多种其他肽。 然而，令人惊讶的是，发现PACAP 1-38和PACAP 6-38是CART结合的低亲和力抑制剂。 CART处理增加了35S [Gamma] GTP-S与PC12细胞膜的结合。此外，完整的PC12细胞的CART处理以通过分化增加的方式引起了磷酸化的稳健增加，并被百日咳毒素阻塞并被PACAP 6-38拮抗。 这些发现表明，PC12细胞中的CART结合位点反映了与GI/O相关的G蛋白偶联受体，并且还证明了PACAP 6-38可以用作CART受体拮抗剂。