METAL CHELATION IN PROTEINS WITH POLYMETALLIC CLUSTERS

蛋白质与多金属簇的金属螯合

基本信息

批准号：
7422317
负责人：
Dennis R. Winge
金额：
$ 48.64万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-05-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7422317
关键词：
ATP phosphohydrolase Abbreviations Acute Address Affect Affinity Alzheimer&apos s Disease Amides Amino Acid Sequence Anabolism Area Attenuated Bacterial Genes Binding Binding Proteins Binding Sites Biochemical Biology Bos taurus Brain Buffers Candida glabrata Cattle Cell Nucleus Cell membrane Cell physiology Cells Ceramides Ceruloplasmin Characteristics Chemistry Class Cleaved cell Codon Nucleotides Complex Condition Consensus Sequence Copper Crystallography Cuprozinc Superoxide Dismutase Cysteine Cytochrome-c Oxidase Deficiency Cytosol DNA DNA Binding DNA Binding Domain DNA Microarray Chip DNA Microarray format DNA-Directed RNA Polymerase Defect Dendrites Depth Distal Docking Elements Employee Strikes Ensure Enzymes Equilibrium Exhibits Figs - dietary Fungal Genome GCG gene Gene Activation Gene Expression Genes Genetic Transcription Glean Glucagon Goals Golgi Apparatus HMGA1a Protein Heme Hepatolenticular Degeneration Homeostasis Homologous Gene Human Hydrogen Hydrogen Bonding Hydroxylation In Vitro Inositol Intestines Ion Transport Ions LIM Domain Protein Ligands Ligation Lobe Localized Macrophage-1 Antigen Major Groove Maps Mediating Membrane Membrane Proteins Metabolism Metal Ion Binding Metallothionein Metals Minor Minor Groove Mitochondria Mitochondrial Proteins Mitolactol Modeling Molecular Molecular Chaperones Molecular Weight Mutation N-terminal NMR Spectroscopy Names Nature Neurofibrillary Tangles Neurons Nuclear Numbers Object Attachment Open Reading Frames Organelles Orthologous Gene Oxidases Pathway interactions Peptide Sequence Determination Peptides Phenotype Phospholipids Physiological Podospora Podospora anserina Positioning Attribute Principal Investigator Process Property Protein Conformation Protein Family Protein Overexpression Proteins Range Reaction Reading Regulation Resolution Respiration Risk Role Route Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Series Side Signal Transduction Site Solutions Specificity Structure Sulfur Superoxide Dismutase Surface System Tertiary Protein Structure Testing Transactivation Transcriptional Activation Vesicle Work Yeasts activating transcription factor base brain tissue cadmium ion chelation cofactor conformer copper(I)-thiolate cupric sulfide cysteine rich protein cytochrome c oxidase dalton in vivo insight member metalloenzyme metallothionein III mitochondrion intermembrane space mutant novel permease polypeptide programs promoter protein function protein protein interaction protein structure protein structure function research study respiratory stoichiometry trafficking transcription factor uptake

项目摘要

The focus is on cysteine-rich proteins that form metal thiolate polymetallic clusters. A paradigm of this class is metallothionein (MT). Polymetallic clusters with distinct properties are induced by Zn (II) and Cu(I) ions. One goal is to determine the magnitude of structural reorganization in MT depending on the type of cluster formed. A second objective is to determine whether similar cluster structure alter the tertiary fold and function of these proteins. Three classes of molecules will be studied. First, a novel metallothionein implicated in Alzheimer's disease will be investigated. The MT, designated as GIF for Growth Inhibitory Factor, is active in inhibiting dendrite formation in neurons induced by Alzheimer's brain extracts. The tangled outgrowths of neurons that is characteristic of Alzheimer's disease may relate to the low concentration of GIF in Alzheimer's brain tissue. We propose experiments to determine which metallo-conformer of GIF is active in reversing the Alzheimer's extract induced proliferation of neurons. A series of experiments are proposed to map the segment of GIF responsible for activity. We plan to characterize the metal clusters in GIF to determine whether sequence differences in MT and GIF affect properties of the polymetallic clusters. The second class of proteins includes two fungal transcription factors. ACE and AMT1. Cu(I) binding to ACE! and AMT1 activates the factors for transcriptional activation of MT genes in Saccharomyces cerevisiae and Candida glabrata, respectively. We propose to characterize the Cu(I) thiolate polymetallic clusters in these two protein conformations. DNA binding sites of CuACE1 and CuAMT1 will be characterized with the goal of elucidating the structure of the transcriptionally active CuAMT1/DNA complex. The third class is the cysteine-rich sequence motif, designated LIM. The metal centers in two LIM-domain proteins, designated Cysteine-Rich Protein (CRP) and Cysteine- Rich Intestinal Protein (CRIP) will be studied to determine whether LIM proteins exhibit metal-induced conformational dynamics. A central postulate is that the structure and function of these classes of proteins are affected by the coordination chemistry of the metal centers. We eventually want to determine the role of specific metal ion binding in function. Molecules in these three classes exhibit a wide range of physiological functions from regulation of DNA transcription (ACE1 and AMT1), metal ion buffering (MT), inhibition of neuron outgrowth (GIF), protein-protein interaction (CRP) and perhaps metal transport (CRIP).

重点是形成金属硫醇盐的富含半胱氨酸的蛋白质多金属簇。此类的一个范例是金属硫蛋白（MT）。具有独特性质的多金属簇是由 Zn (II) 和 Cu(I) 离子。目标之一是确定结构性影响的大小 MT 中的重组取决于形成的簇的类型。一秒钟目的是确定相似的簇结构是否会改变这些蛋白质的三级折叠和功能。三类分子将被研究。首先，一种新型金属硫蛋白涉及阿尔茨海默病将得到研究。 MT，指定为 GIF 生长抑制因子，具有抑制树突形成的活性阿尔茨海默氏症大脑提取物诱导的神经元。纠结的产物阿尔茨海默病特征性的神经元数量可能与阿尔茨海默病脑组织中 GIF 浓度低。我们建议确定 GIF 的哪种金属构象在其中具有活性的实验逆转阿尔茨海默病提取物诱导的神经元增殖。一个提出了一系列实验来映射 GIF 负责的部分用于活动。我们计划表征 GIF 中的金属簇确定 MT 和 GIF 中的序列差异是否影响属性的多金属簇。第二类蛋白质包括两种真菌转录因子。 ACE 和 AMT1。 Cu(I) 与 ACE 结合！和 AMT1 激活 MT 基因转录激活因子分别是酿酒酵母和光滑假丝酵母。我们建议表征这两个化合物中的 Cu(I) 硫醇盐多金属簇蛋白质构象。 CuACE1 和 CuAMT1 的 DNA 结合位点将是其特征在于阐明结构的目标具有转录活性的 CuAMT1/DNA 复合物。第三类是富含半胱氨酸的序列基序，称为 LIM。金属中心分为两部分 LIM 结构域蛋白，称为富含半胱氨酸蛋白 (CRP) 和半胱氨酸- 将研究丰富肠蛋白 (CRIP) 以确定 LIM 是否蛋白质表现出金属诱导的构象动力学。一个中央假设这些类别的蛋白质的结构和功能受金属中心配位化学的影响。我们最终想要确定特定金属离子结合的作用功能。这三类分子表现出广泛的 DNA 转录调节（ACE1 和 AMT1）、金属离子缓冲（MT）、神经元生长抑制（GIF）、蛋白质-蛋白质相互作用（CRP），或许还有金属转运（CRIP）。