2A29

The solution structure of the AMP-PNP bound nucleotide binding domain of KdpB

「1X6K」から置き換えられました

2A29 の概要

• エントリーDOI: 10.2210/pdb2a29/pdb
• 関連するPDBエントリー: 1SVJ 1U7Q 2A00
• NMR情報: BMRB: 6029,6030
• 分子名称: Potassium-transporting ATPase B chain, PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER (2 entities in total)
• 機能のキーワード: alpha-beta sandwich, hydrolase
• 由来する生物種: Escherichia coli
• 細胞内の位置: Cell inner membrane; Multi-pass membrane protein: P03960
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 17674.49

構造登録者

Haupt, M.,Bramkamp, M.,Coles, M.,Altendorf, K.,Kessler, H. (登録日: 2005-06-22, 公開日: 2005-12-20, 最終更新日: 2024-05-29)

主引用文献

Haupt, M.,Bramkamp, M.,Heller, M.,Coles, M.,Deckers-Hebestreit, G.,Herkenhoff-Hesselmann, B.,Altendorf, K.,Kessler, H.
The Holo-form of the Nucleotide Binding Domain of the KdpFABC Complex from Escherichia coli Reveals a New Binding Mode
J.Biol.Chem., 281:9641-9649, 2006

PubMed Abstract: P-type ATPases are ubiquitously abundant enzymes involved in active transport of charged residues across biological membranes. The KdpB subunit of the prokaryotic Kdp-ATPase (KdpFABC complex) shares characteristic regions of homology with class II-IV P-type ATPases and has been shown previously to be misgrouped as a class IA P-type ATPase. Here, we present the NMR structure of the AMP-PNP-bound nucleotide binding domain KdpBN of the Escherichia coli Kdp-ATPase at high resolution. The aromatic moiety of the nucleotide is clipped into the binding pocket by Phe(377) and Lys(395) via a pi-pi stacking and a cation-pi interaction, respectively. Charged residues at the outer rim of the binding pocket (Arg(317), Arg(382), Asp(399), and Glu(348)) stabilize and direct the triphosphate group via electrostatic attraction and repulsion toward the phosphorylation domain. The nucleotide binding mode was corroborated by the replacement of critical residues. The conservative mutation F377Y produced a high residual nucleotide binding capacity, whereas replacement by alanine resulted in low nucleotide binding capacities and a considerable loss of ATPase activity. Similarly, mutation K395A resulted in loss of ATPase activity and nucleotide binding affinity, even though the protein was properly folded. We present a schematic model of the nucleotide binding mode that allows for both high selectivity and a low nucleotide binding constant, necessary for the fast and effective turnover rate realized in the reaction cycle of the Kdp-ATPase.

PubMed: 16354672
DOI: 10.1074/jbc.M508290200

実験手法

SOLUTION NMR