1E5H

DELTA-R307A DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH SUCCINATE AND CARBON DIOXIDE

1E5H の概要

• エントリーDOI: 10.2210/pdb1e5h/pdb
• 関連するPDBエントリー: 1DCS 1E5I 1RXF 1RXG
• 分子名称: DEACETOXYCEPHALOSPORIN C SYNTHASE, FE (II) ION, SUCCINIC ACID, ... (5 entities in total)
• 機能のキーワード: ferrous oxygenase, cephalosporin, 2-oxoglutarate, c-terminus antibiotics, oxidoreductase, oxidative coupling control
• 由来する生物種: STREPTOMYCES CLAVULIGERUS
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34492.22

構造登録者

Lee, H.J.,Lloyd, M.D.,Harlos, K.,Clifton, I.J.,Baldwin, J.E.,Schofield, C.J. (登録日: 2000-07-26, 公開日: 2001-07-26, 最終更新日: 2023-12-13)

主引用文献

Lee, H.J.,Lloyd, M.D.,Harlos, K.,Clifton, I.J.,Baldwin, J.E.,Schofield, C.J.
Kinetic and Crystallographic Studies on Deacetoxycephalosporin C Synthase (Daocs)
J.Mol.Biol., 308:937-, 2001

PubMed Abstract: Deacetoxycephalosporin C synthase (DAOCS) is an iron(II) and 2-oxoglutarate-dependent oxygenase that catalyzes the conversion of penicillin N to deacetoxycephalosporin C, the committed step in the biosynthesis of cephalosporin antibiotics. The crystal structure of DAOCS revealed that the C terminus of one molecule is inserted into the active site of its neighbor in a cyclical fashion within a trimeric unit. This arrangement has hindered the generation of crystalline enzyme-substrate complexes. Therefore, we constructed a series of DAOCS mutants with modified C termini. Oxidation of 2-oxoglutarate was significantly uncoupled from oxidation of the penicillin substrate in certain truncated mutants. The extent of uncoupling varied with the number of residues deleted and the penicillin substrate used. Crystal structures were determined for the DeltaR306 mutant complexed with iron(II) and 2-oxoglutarate (to 2.10 A) and the DeltaR306A mutant complexed with iron(II), succinate and unhydrated carbon dioxide (to 1.96 A). The latter may mimic a product complex, and supports proposals for a metal-bound CO(2) intermediate during catalysis.

PubMed: 11352583
DOI: 10.1006/JMBI.2001.4649

実験手法

X-RAY DIFFRACTION (1.96 Å)