1TXE

Solution structure of the active-centre mutant Ile14Ala of the histidine-containing phosphocarrier protein (HPr) from Staphylococcus carnosus

1TXE の概要

• エントリーDOI: 10.2210/pdb1txe/pdb
• 関連するPDBエントリー: 1QR5
• NMR情報: BMRB: 6254
• 分子名称: Phosphocarrier protein HPr (1 entity in total)
• 機能のキーワード: open-faced beta-sandwich, structural proteomics in europe, spine, structural genomics, transport protein
• 由来する生物種: Staphylococcus carnosus
• 細胞内の位置: Cytoplasm: P23534
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 9476.57

構造登録者

Moeglich, A.,Koch, B.,Hengstenberg, W.,Brunner, E.,Kalbitzer, H.R.,Structural Proteomics in Europe (SPINE) (登録日: 2004-07-04, 公開日: 2005-03-08, 最終更新日: 2024-05-29)

主引用文献

Moeglich, A.,Koch, B.,Gronwald, W.,Hengstenberg, W.,Brunner, E.,Kalbitzer, H.R.
Solution structure of the active-centre mutant I14A of the histidine-containing phosphocarrier protein from Staphylococcus carnosus
Eur.J.Biochem., 271:4815-4824, 2004

PubMed Abstract: High-pressure NMR experiments performed on the histidine-containing phosphocarrier protein (HPr) from Staphylococcus carnosus have shown that residue Ile14, which is located in the active-centre loop, exhibits a peculiarly small pressure response. In contrast, the rest of the loop shows strong pressure effects as is expected for typical protein interaction sites. To elucidate the structural role of this residue, the mutant protein HPr(I14A), in which Ile14 is replaced by Ala, was produced and studied by solution NMR spectroscopy. On the basis of 1406 structural restraints including 20 directly detected hydrogen bonds, 49 1H(N)-15N, and 25 1H(N)-1Halpha residual dipolar couplings, a well resolved three-dimensional structure could be determined. The overall fold of the protein is not influenced by the mutation but characteristic conformational changes are introduced into the active-centre loop. They lead to a displacement of the ring system of His15 and a distortion of the N-terminus of the first helix, which supports the histidine ring. In addition, the C-terminal helix is bent because the side chain of Leu86 located at the end of this helix partly fills the hydrophobic cavity created by the mutation. Xenon, which is known to occupy hydrophobic cavities, causes a partial reversal of the mutation-induced structural effects. The observed structural changes explain the reduced phosphocarrier activity of the mutant and agree well with the earlier suggestion that Ile14 represents an anchoring point stabilizing the active-centre loop in its correct conformation.

PubMed: 15606769
DOI: 10.1111/j.1432-1033.2004.04447.x

実験手法

SOLUTION NMR