3M0V

Crystal structure of Pseudomonas stutzeri L-rhamnose isomerase mutant S329L in complex with L-rhamnose

3M0V の概要

• エントリーDOI: 10.2210/pdb3m0v/pdb
• 関連するPDBエントリー: 2HCV 2I56 2I57 3M0H 3M0L 3M0M 3M0X 3M0Y
• 分子名称: L-rhamnose isomerase, MANGANESE (II) ION, L-RHAMNOSE, ... (4 entities in total)
• 機能のキーワード: beta/alpha barrel, homo-tetramer, metal-binding protein, tim barrel, isomerase
• 由来する生物種: Pseudomonas stutzeri (Pseudomonas perfectomarina)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 193218.89

構造登録者

Yoshida, H.,Takeda, K.,Izumori, K.,Kamitori, S. (登録日: 2010-03-03, 公開日: 2010-11-10, 最終更新日: 2023-11-01)

主引用文献

Yoshida, H.,Takeda, K.,Izumori, K.,Kamitori, S.
Elucidation of the role of Ser329 and the C-terminal region in the catalytic activity of Pseudomonas stutzeri L-rhamnose isomerase
Protein Eng.Des.Sel., 23:919-927, 2010

PubMed Abstract: Pseudomonas stutzeri l-rhamnose isomerase (l-RhI) is capable of catalyzing the isomerization between various aldoses and ketoses, showing high catalytic activity with broad substrate-specificity compared with Escherichia coli l-RhI. In a previous study, the crystal structure of P. stutzeri l-RhI revealed an active site comparable with that of E. coli l-RhI and d-xylose isomerases (d-XIs) with structurally conserved amino acids, but also with a different residue seemingly responsible for the specificity of P. stutzeri l-RhI, though the residue itself does not interact with the bound substrate. This residue, Ser329, corresponds to Phe336 in E. coli l-RhI and Lys294 in Actinoplanes missouriensis d-XI. To elucidate the role of Ser329 in P. stutzeri l-RhI, we constructed mutants, S329F (E. coli l-RhI type), S329K (A. missouriensis d-XI type), S329L and S329A. Analyses of the catalytic activity and crystal structure of the mutants revealed a hydroxyl group of Ser329 to be crucial for catalytic activity via interaction with a water molecule. In addition, in complexes with substrate, the mutants S329F and S329L exhibited significant electron density in the C-terminal region not observed in the wild-type P. stutzeri l-RhI. The C-terminal region of P. stutzeri l-RhI has flexibility and shows a flip-flop movement at the inter-molecular surface of the dimeric form.

PubMed: 20977999
DOI: 10.1093/protein/gzq077

実験手法

X-RAY DIFFRACTION (1.79 Å)