CHAPERONE TARGET INTERACTIONS

陪伴目标互动

• 批准号: 7690594
• 负责人: Ninian J Blackburn
• 金额: $ 28.86万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7690594
• 关键词: ATP phosphohydrolase; Active Sites; Affinity; Antioxidants; Ataxia; Binding; Binding Sites; Biogenic Amine Metabolism; C-terminal; Carrier Proteins; Cells; Chemistry; Collaborations; Comparative Study; Complex; Connective Tissue; Copper; Coupled; Cysteine; Defect; Development; Disease; Disulfides; Endocrine Gland Neoplasms; Enzymes; Eukaryota; Failure; Funding; Gene Mutation; Genes; Grant; Hand; Health; Hepatolenticular Degeneration; Homeostasis; Human; Indium; Ions; Iron; Label; Laboratories; Ligation; Link; Malignant Neoplasms; Measures; Mediating; Membrane; Menkes Kinky Hair Syndrome; Metals; Methodology; Methods; Molecular; Molecular Chaperones; Monitor; Mutagenesis; Neurodegenerative Disorders; Neuropeptides; Neurotransmitters; Organism; Oxidation-Reduction; Pathology; Pathway interactions; Peptides; Pigmentation physiologic function; Plasmids; Play; Process; Proteins; Recruitment Activity; Regulation; Respiration; Roentgen Rays; Role; Route; Secretory Vesicles; Selenium; Selenocysteine; Selenomethionine; Senile Plaques; Side; Site; Spectrum Analysis; Structure; System; Techniques; Technology; Testing; Wilson disease protein; absorption; base; cancer therapy; cancer type; copper-transporting ATPase; design; enzyme activity; hormone biosynthesis; innovation; intermolecular interaction; meetings; mutant; novel; orexin A receptor; peptidylglycine alpha-amidating monooxygenase; permease; reconstitution; stoichiometry; structural biology; synthetic peptide; trafficking; tumor growth; uptake

Copper is an essential element in eukaryotic organisms. Due to facile redox chemistry at moderate potentials, it functions at the active sites of many critical enzymes involved in a wide variety of processes, including neurotransmitter and neuropeptide hormone biosynthesis, biogenic amine metabolism and connective tissue formation, pigmentation, respiration, and antioxidant defense. New findings have linked certain cancers to copper mediated enzyme activity, for example the observation that the expression of the cuproenzyme PAM in endocrine tumors mediates tumor growth. Another intriguing link of importance to cancer therapy, is the recruitment of the CTR1 high affinity copper transporter in the uptake of cis-platin into cells by a presently unknown mechanism. Because copper is both essential and potentially toxic, cells have evolved complex molecular machinery to maintain levels within a narrow range of concentration, and failure to achieve liomeostasis results in diseases related to either Cu deficiency or excess which include those involving genetic mutations in the copper transporting ATPases, ATP7A (Menkes disease) and ATP7B (Wilson disease). Important consequences of aberrant copper homeostasis have also been implicated in neurodegenerative disorders, and in the pathology of amyloid plaque formation in Alzheimers. Therefore, the study of copper uptake, transport and trafficking is of great importance to human health. In eukaryotes, copper is imported by the membrane associated high affinity transporter CTR1, and thence distributed to target proteins via metallochaperone shuttles. An important tenet of copper homeostasis is the hypothesis that CTR1 hands off copper to the chaperones which in turn transport the metal to its sites of action. However, the interaction of CTR1 with chaperones has not yet been experimentally verified. Project 2 of the PPG will therefore leverage recent advances in the structural biology of hCTRl to investigate (i) the mode of copper binding to CTR1, (ii) the interaction of CTR1 with the chaperone hCCS, and (iii) the mechanisms by which chaperones and ATPases interact with and transfer copper to their target cuproenzymes. The technique of selenium-labeling coupled to X-ray absorption spectroscopy (developed in the last grant period) will be used extensively in these studies.

铜是真核生物的必需元素。由于在中等电位下容易的氧化还原化学， 在许多关键酶的活性位点发挥作用，这些关键酶参与多种过程，包括 神经递质和神经肽激素生物合成、生物胺代谢和结缔组织 形成，色素沉着，呼吸和抗氧化防御。新的发现将某些癌症与 铜介导的酶活性，例如观察到铜酶PAM在细胞中的表达， 内分泌肿瘤介导肿瘤生长。另一个有趣的联系，重要的癌症治疗，是 本发明涉及通过目前的免疫调节剂在顺铂摄取到细胞中的CTR 1高亲和力铜转运蛋白的募集。 未知机制由于铜是必需的，也有潜在的毒性，细胞已经进化出复杂的 分子机器维持在一个狭窄的浓度范围内的水平，并未能实现 体内平衡导致与铜缺乏或过量相关的疾病，包括那些涉及遗传的疾病。 铜转运ATP酶，ATP 7A（门克斯病）和ATP 7B（威尔逊病）的突变。 异常铜稳态的重要后果也涉及神经退行性疾病。 在阿尔茨海默病中的淀粉样蛋白斑形成的病理学中。因此，对铜的研究 摄取、运输和贩运对人类健康极为重要。在真核生物中，铜是通过 膜结合的高亲和力转运蛋白CTR 1，并由此通过 金属伴侣穿梭。铜稳态的一个重要原则是假设CTR1传递给 分子伴侣又将金属运送到其作用位点。然而， 具有分子伴侣的CTR 1尚未得到实验验证。因此，项目编制小组的项目2将利用 在hCTR 1的结构生物学方面的最新进展，以研究（i）铜与CTR 1结合的模式，（ii） CTR1与分子伴侣hCCS的相互作用，以及（iii）分子伴侣和ATP酶 与铜相互作用并将铜转移到它们的目标铜酶。硒标记技术与 在这些研究中将广泛使用X射线吸收光谱法（在上一个资助期开发）。