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发生氧化还原化学反应， 在涉及多种过程的许多关键酶的活性位点发挥功能，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。新发现将某些癌症与 铜介导的酶活性，例如观察到铜酶 PAM 的表达 内分泌肿瘤介导肿瘤生长。癌症治疗的另一个重要的有趣联系是 CTR1高亲和力铜转运蛋白在细胞摄取顺铂过程中的募集 未知的机制。由于铜既是必需的又具有潜在的毒性，因此细胞进化出了复杂的 分子机械将水平维持在狭窄的浓度范围内，并且未能实现 脂质平衡会导致与铜缺乏或过量相关的疾病，其中包括那些涉及遗传的疾病 铜转运 ATP 酶、ATP7A（门克斯病）和 ATP7B（威尔逊病）发生突变。 铜稳态异常的重要后果也与神经退行性疾病有关 疾病，以及阿尔茨海默病中淀粉样蛋白斑形成的病理学。因此，铜的研究 摄取、运输和贩运对人类健康非常重要。在真核生物中，铜的输入方式是 膜相关的高亲和力转运蛋白 CTR1，然后通过 金属伴侣穿梭。铜稳态的一个重要原则是 CTR1 传递的假设 将铜结合到分子伴侣上，分子伴侣又将金属转运到其作用位点。然而，相互作用 带有伴侣的 CTR1 尚未经过实验验证。因此，PPG 的项目 2 将利用 hCTR1 结构生物学的最新进展，以研究 (i) 铜与 CTR1 结合的模式，(ii) CTR1 与伴侣 hCCS 的相互作用，以及 (iii) 伴侣和 ATP 酶的机制 与目标铜酶相互作用并将铜转移至目标铜酶。硒标记技术与 X 射线吸收光谱（在最后资助期间开发）将广泛用于这些研究。