1KS9

Ketopantoate Reductase from Escherichia coli

1KS9 の概要

• エントリーDOI: 10.2210/pdb1ks9/pdb
• 分子名称: 2-DEHYDROPANTOATE 2-REDUCTASE (2 entities in total)
• 機能のキーワード: pane, apba, ketopantoate reductase, rossmann fold, monomer, apo, oxidoreductase
• 由来する生物種: Escherichia coli
• 細胞内の位置: Cytoplasm : P0A9J4
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 32479.06

構造登録者

Matak-Vinkovic, D.,Vinkovic, M.,Saldanha, S.A.,Ashurst, J.A.,von Delft, F.,Inoue, T.,Miguel, R.N.,Smith, A.G.,Blundell, T.L.,Abell, C. (登録日: 2002-01-11, 公開日: 2002-01-25, 最終更新日: 2024-11-13)

主引用文献

Matak-Vinkovic, D.,Vinkovic, M.,Saldanha, S.A.,Ashurst, J.L.,von Delft, F.,Inoue, T.,Miguel, R.N.,Smith, A.G.,Blundell, T.L.,Abell, C.
Crystal structure of Escherichia coli ketopantoate reductase at 1.7 A resolution and insight into the enzyme mechanism.
Biochemistry, 40:14493-14500, 2001

PubMed Abstract: Ketopantoate reductase (KPR, EC 1.1.1.169) catalyzes the NADPH-dependent reduction of ketopantoate to pantoate on the pantothenate (vitamin B(5)) biosynthetic pathway. The Escherichia coli panE gene encoding KPR was cloned and expressed at high levels as the native and selenomethionine-substituted (SeMet) proteins. Both native and SeMet recombinant proteins were purified by three chromatographic steps, to yield pure proteins. The wild-type enzyme was found to have a K(M)(NADPH) of 20 microM, a K(M)(ketopantoate) of 60 microM, and a k(cat) of 40 s(-1). Regular prismatic KPR crystals were prepared using the hanging drop technique. They belonged to the tetragonal space group P4(2)2(1)2, with cell parameters: a = b = 103.7 A and c = 55.7 A, accommodating one enzyme molecule per asymmetric unit. The structure of KPR was determined by the multiwavelength anomalous dispersion method using the SeMet protein, for which data were collected to 2.3 A resolution. The native data were collected to 1.7 A resolution and used to refine the final structure. The secondary structure comprises 12 alpha-helices, three 3(10)-helices, and 11 beta-strands. The enzyme is monomeric and has two domains separated by a cleft. The N-terminal domain has an alphabeta-fold of the Rossmann type. The C-terminal domain (residues 170-291) is composed of eight alpha-helices. KPR is shown to be a member of the 6-phosphogluconate dehydrogenase C-terminal domain-like superfamily. A model for the ternary enzyme-NADPH-ketopantoate ternary complex provides a rationale for kinetic data reported for specific site-directed mutants.

PubMed: 11724562
DOI: 10.1021/bi011020w

実験手法

X-RAY DIFFRACTION (1.7 Å)