3ST7

Crystal Structure of capsular polysaccharide assembling protein CapF from staphylococcus aureus

3ST7 の概要

• エントリーDOI: 10.2210/pdb3st7/pdb
• 関連するPDBエントリー: 2ZKL
• 分子名称: Capsular polysaccharide synthesis enzyme Cap5F, ZINC ION, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: rossmann fold, cupid domain, short-chain dehydrogenase/reductase, nadph, udp-4-hexulose reductase, oxidoreductase
• 由来する生物種: Staphylococcus aureus
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 42814.69

構造登録者

Miyafusa, T.,Tanaka, Y.,Kuroda, M.,Yao, M.,Watanabe, M.,Ohta, T.,Tanaka, I.,Caaveiro, J.M.M.,Tsumoto, K. (登録日: 2011-07-09, 公開日: 2012-02-15, 最終更新日: 2024-11-20)

主引用文献

Miyafusa, T.,Caaveiro, J.M.,Tanaka, Y.,Tsumoto, K.
Crystal structure of the enzyme CapF of Staphylococcus aureus reveals a unique architecture composed of two functional domains.
Biochem.J., 443:671-680, 2012

PubMed Abstract: CP (capsular polysaccharide) is an important virulence factor during infections by the bacterium Staphylococcus aureus. The enzyme CapF is an attractive therapeutic candidate belonging to the biosynthetic route of CP of pathogenic strains of S. aureus. In the present study, we report two independent crystal structures of CapF in an open form of the apoenzyme. CapF is a homodimer displaying a characteristic dumb-bell-shaped architecture composed of two domains. The N-terminal domain (residues 1-252) adopts a Rossmann fold belonging to the short-chain dehydrogenase/reductase family of proteins. The C-terminal domain (residues 252-369) displays a standard cupin fold with a Zn2+ ion bound deep in the binding pocket of the β-barrel. Functional and thermodynamic analyses indicated that each domain catalyses separate enzymatic reactions. The cupin domain is necessary for the C3-epimerization of UDP-4-hexulose. Meanwhile, the N-terminal domain catalyses the NADPH-dependent reduction of the intermediate species generated by the cupin domain. Analysis by ITC (isothermal titration calorimetry) revealed a fascinating thermodynamic switch governing the attachment and release of the coenzyme NADPH during each catalytic cycle. These observations suggested that the binding of coenzyme to CapF facilitates a disorder-to-order transition in the catalytic loop of the reductase (N-terminal) domain. We anticipate that the present study will improve the general understanding of the synthesis of CP in S. aureus and will aid in the design of new therapeutic agents against this pathogenic bacterium.

PubMed: 22320426
DOI: 10.1042/BJ20112049

実験手法

X-RAY DIFFRACTION (2.45 Å)