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

描述（由申请人提供）：维持金属稳态对正常代谢功能至关重要，体内金属的流入/排出必须严格管理。控制金属运输的一种方式是通过p1b型atp酶，这是一个整合膜蛋白的超家族，将atp水解与跨膜过渡金属阳离子运输结合起来。关于p1b -5- atp酶还有很多未知的地方，包括金属底物的身份和酶的c端氰氰酯（Hr）样结构域的功能。本研究旨在利用光谱和结构技术，通过探索C端双铁中心与生理相关气体O2和NO的相互作用，来阐明h -like结构域的功能。此外，本提案旨在确定结合在P1B-5- atp酶跨膜区域的天然金属底物的身份。为了探索金属底物与P1B-5 atp酶多肽之间相互作用的性质，将进行光谱和结构工作。这些研究将有助于阐明p1b -5- atp酶家族中Hr-like结构域的功能。此外，这项工作将促进对p1b -5- atp酶在过渡金属运输中的功能的理解。

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.