Neural EP24.15- A Model for Neuropeptidase Function

神经 EP24.15- 神经肽酶功能模型

• 批准号: 7151912
• 负责人: Marc J Glucksman
• 金额: $ 33.21万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-20 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7151912
• 关键词: Affinity; Alzheimer' s Disease; Antibodies; Binding; Binding Sites; Biochemical Genetics; Biological; Biological Assay; Brain; Cancer Cell Growth; Catalysis; Cells; Class; Cleaved cell; Collaborations; Complementary DNA; Complex; Custom; Cytosol; Development; Endocrine; Environment; Enzyme Kinetics; Enzymes; Eukaryotic Cell; Family; Gel; Goals; Growth and Development function; Hand; Immune response; Kinetics; Laboratories; Left; Ligands; Major Histocompatibility Complex; Malignant Neoplasms; Malignant neoplasm of prostate; Mass Spectrum Analysis; Metalloendopeptidases; Modeling; Modification; Molecular Biology; Movement; Neurodegenerative Disorders; Neuropeptides; Neurosecretory Systems; Organ; Pain; Peptides; Perception; Phage Display; Pharmaceutical Preparations; Pituitary Gland; Play; Production; Protein Binding; Protein Chemistry; Protein Isoforms; Proteins; Proteomics; Rate; Reagent; Regulation; Research Personnel; Role; Site; Site-Directed Mutagenesis; Specificity; Structural Models; Testing; Two-Dimensional Gel Electrophoresis; Upper arm; Viral; Work; X ray diffraction analysis; X-Ray Diffraction; design; extracellular; hormone regulation; inhibitor/antagonist; macromolecule; malignant endocrine gland neoplasm; member; molecular modeling; mutant; novel therapeutics; peptide I; programs; relating to nervous system; reproductive; size; small molecule; structural biology

DESCRIPTION (provided by applicant): 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 understanding the 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.

描述（由申请人提供）：本提案旨在研究E.C.3.4.24.15（EP24.15）的底物结合、亚型、相关蛋白和底物大小特异性，目的是利用新治疗药物和小分子的设计，以调节神经内分泌/神经退行性疾病和内分泌癌的发展。EP 24.15是金属内肽酶家族的原型成员，广泛表达于脑、垂体和生殖器官。在细胞外，EP 24.15切割和调节神经肽，因此EP 24.15在神经内分泌激素调节、疼痛感知、神经退行性疾病的发展和前列腺癌细胞生长中起重要作用。本提案的目的是进一步了解EP 24.15的功能，其亚型，以及密切相关的金属内肽酶，目的是了解EP 24.15在神经内分泌/神经退行性疾病和癌症中的突出作用。因此，该提案将整合生物化学，遗传学，结构和蛋白质组学方法来研究EP 24.15的底物结合，异构体，相关蛋白和底物大小特异性。因此，具体目标是：具体目标1。阐明赋予底物结合和催化作用的功能位点。瞄准洛杉矶。确定哪些残基构成EP 24.15的底物（抑制剂）结合位点。瞄准Ib。确定哪些残基对底物催化和特异性至关重要。具体目标2.确定EP 24.15和相关酶的亚型和蛋白质结合伴侣。目标2a。鉴定EP 24.15和EP 24.16的细胞外和细胞内亚型。目标2b。表征EP 24.15与相关蛋白结合伴侣的复合物。具体目标3.揭示新的基底和基底尺寸选择性的机理。目标3a。确定EP 24.15存在哪些其他（神经）肽底物。目标3b。阐明底物结合后的结构变化，重点是结构域的运动。