Neural EP24.15- A Model for Neuropeptidase Function

这项建议的目的是研究底物结合，亚型，相关蛋白， 和E.C.3.4.24.15(EP24.15)的底物大小特异性，旨在为新的治疗方法的设计提供帮助 药物和小分子，调节神经内分泌/神经退行性疾病的发展和 内分泌癌(S)。 EP24.15是金属内肽酶家族的原型成员，广泛表达于 大脑、脑下垂体和生殖器官。在细胞外，EP24.15裂解和调节神经肽，从而 EP24.15在神经内分泌激素调节、痛觉、心力衰竭的发生发展中起重要作用 神经退行性疾病，以及前列腺癌细胞的生长。这项提议的目标是推动 了解EP24.15、其异构体和密切相关的金属内肽酶的功能 目的了解EP24.15在神经内分泌/神经退行性疾病和癌症中的突出作用。 因此，这项建议将结合生化、遗传、结构和蛋白质组学方法进行研究。 EP24.15的底物结合、异构体、相关蛋白和底物大小专一性。 因此，具体目标是： 特异目的1.阐明与底物结合和催化有关的功能位点。 瞄准啦。确定哪些残基构成EP24.15的底物(抑制物)结合部位。 瞄准Ib.确定哪些残基对底物催化和特异性至关重要。 特异性目的2.确定EP24.15及其相关酶的异构体和蛋白结合伙伴。 目标2a。鉴定EP24.15和EP24.16的胞外和胞内亚型。 目标2b。表征EP24.15与相关蛋白结合伙伴的复合体。 具体目的3.揭示新的底物和底物尺寸选择性的机理。 目标3a。确定EP24.15还存在哪些(神经肽)底物。 目标3b。阐明底物结合时的结构变化，重点是结构域的移动。

The objective of this proposal is to investigate the substrate binding, isoforms, associated proteins, and substrate size specificity of E.C.3.4.24.15 (EP24.15), with the goal of availing design of new therapeutic drugs and small molecules, to modulate development of neuroendocrine/neurodegenerative diseases and endocrine cancer(s). EP24.15 is the prototypic member of the metalloendopeptidase family, and is widely expressed in the brain, pituitary, and reproductive organs. Extracellularly, EP24.15 cleaves and modulates neuropeptides, and thus EP24.15 plays a significant role in neuroendocrine hormone regulation, pain perception, development of neurodegenerative diseases, and prostate cancer cell growth. The goal of this proposal is to further the understanding of the functions of EP24.15, of its isoforms, and of closely related metalloendopeptidases, with the goal of understandingthe prominence of EP24.15 in neuroendocrine/neurodegenerative diseases and cancer. Therefore, this proposal will integrate biochemical, genetic, structural, and proteomic approaches to study substrate binding, isoforms, associated proteins, and substrate size specificity of EP24.15. Therefore, the specific aims are: SPECIFIC AIM 1. Elucidate functional sites conferring substrate binding and catalysis. Aim la. Determine which residues comprise the substrate (inhibitor) binding site of EP24.15. Aim Ib. Determine which residues are crucial for substrate catalysis and specificity. SPECIFIC AIM 2. Determine isoforms and protein binding partners of EP24.15 and of related enzymes. Aim 2a. Identify the extracellular and intracellular isoforms of EP24.15 and EP24.16. Aim 2b. Characterize complexes of EP24.15 with associated protein-binding partners. SPECIFIC AIM 3. Unveil new substrates and mechanism of substrate size selectivity. Aim 3a. Determine what other (neuro)peptide substrates exist for EP24.15. Aim 3b. Elucidate structural changes upon substrate binding, focusing on domain movements.