Characterization of the P1B-5ATPase Hemerythrin-like and Metal-Binding Domains

P1B-5ATP 酶血红蛋白样和金属结合结构域的表征

• 批准号: 8868135
• 负责人: Aaron T Smith
• 金额: $ 5.6万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8868135
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Affect; Amino Acids; Azides; Binding; Binding Sites; Biochemical; Biological Assay; C-terminal; Cations; Crystallization; Data; Enzymes; Family; Gases; Health; Hemerythrin; Hepatolenticular Degeneration; Homeostasis; Human; Integral Membrane Protein; Life; Link; Melilotus; Membrane; Menkes Kinky Hair Syndrome; Metabolic; Metal Binding Site; Metals; Molecular; Mutagenesis; Mutation; N-terminal; Nature; Optics; Organism; Play; Proteins; Pyruvate Kinase; Role; Sequence Analysis; Structure; Techniques; Tertiary Protein Structure; Transition Elements; Transmembrane Domain; Work; adduct; base; in vivo; inorganic phosphate; insight; novel; polypeptide; screening; spectroscopic survey; stoichiometry; three dimensional structure

DESCRIPTION (provided by applicant): Maintaining metal homeostasis is essential for normal metabolic functions, and metal influx/egress must be tightly managed in vivo. One manner in which metal transport is controlled is via the P1B-type ATPases, which are a superfamily of integral membrane proteins that couple ATP-hydrolysis to transmembrane transition metal cation transport. Much is unknown about the P1B-5-ATPase, including the identity of the metal substrate(s) and the function of the enzyme's C-terminal hemerythrin(Hr)-like domain. This proposal aims to use spectroscopic and structural techniques to elucidate the function of the Hr-like domain by exploring the interaction of the C- terminal diiron center with the physiologically relevant gases O2 and NO. Furthermore, this proposal seeks to establish the identity of the native metal substrate(s) that binds in the transmembrane region of the P1B-5- ATPase. Spectroscopic and structural work will be undertaken in order to probe the nature of interaction between the metal substrate(s) and the P1B-5 ATPase polypeptide. These studies will help elucidate the function of the Hr-like domain in the P1B-5-ATPase family. Moreover, this work will advance the understanding of the function of the P1B-5-ATPase in transition metal transport.

描述（由申请人提供）：维持金属稳态对于正常代谢功能至关重要，必须在体内严格管理金属流入/流出。控制金属转运的一种方式是通过P1 B型ATP酶，其是将ATP水解与跨膜过渡金属阳离子转运偶联的整合膜蛋白超家族。关于P1 B-5-ATP酶，包括金属底物的身份和酶的C-末端血红蛋白（Hr）样结构域的功能还有很多未知。该提案旨在使用光谱和结构技术，通过探索C-末端二铁中心与生理相关气体O2和NO的相互作用来阐明Hr样结构域的功能。此外，该提案旨在确定在P1 B-5-ATP酶的跨膜区域中结合的天然金属底物的身份。将进行光谱和结构工作，以探索金属基底和P1 B-5 ATP酶多肽之间相互作用的性质。这些研究将有助于阐明类Hr结构域在P1 B-5-ATP酶家族中的功能。此外，这项工作将进一步了解P1 B-5-ATP酶在过渡金属转运中的功能。