1UPC

Carboxyethylarginine synthase from Streptomyces clavuligerus

1UPC の概要

• エントリーDOI: 10.2210/pdb1upc/pdb
• 関連するPDBエントリー: 1UPA 1UPB
• 分子名称: CARBOXYETHYLARGININE SYNTHASE, THIAMINE DIPHOSPHATE, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: synthase, clavulanic acid, antibiotic, lactamase, flavoprotein, thiamine pyrophosphate
• 由来する生物種: STREPTOMYCES CLAVULIGERUS
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 369639.44

構造登録者

Caines, M.E.C.,Elkins, J.M.,Hewitson, K.S.,Schofield, C.J. (登録日: 2003-09-29, 公開日: 2003-11-20, 最終更新日: 2024-05-08)

主引用文献

Caines, M.E.C.,Elkins, J.M.,Hewitson, K.S.,Schofield, C.J.
Crystal Structure and Mechanistic Implications of N2-(2-Carboxyethyl)Arginine Synthase, the First Enzyme in the Clavulanic Acid Biosynthesis Pathway
J.Biol.Chem., 279:5685-, 2004

PubMed Abstract: The initial step in the biosynthesis of the clinically important beta-lactamase inhibitor clavulanic acid involves condensation of two primary metabolites, D-glyceraldehyde 3-phosphate and L-arginine, to give N2-(2-carboxyethyl)arginine, a beta-amino acid. This unusual N-C bond forming reaction is catalyzed by the thiamin diphosphate (ThP2)-dependent enzyme N2-(2-carboxyethyl)arginine synthase. Here we report the crystal structure of N2-(2-carboxyethyl)arginine synthase, complexed with ThP2 and Mg2+, to 2.35-A resolution. The structure was solved in two space groups, P2(1)2(1)2(1) and P2(1)2(1)2. In both, the enzyme is observed in a tetrameric form, composed of a dimer of two more tightly associated dimers, consistent with both mass spectrometric and gel filtration chromatography studies. Both ThP2 and Mg2+ cofactors are present at the active site, with ThP2 in a "V" conformation as in related enzymes. A sulfate anion is observed in the active site of the enzyme in a location proposed as a binding site for the phosphate group of the d-glyceraldehyde 3-phosphate substrate. The mechanistic implications of the active site arrangement are discussed, including the potential role of the aminopyrimidine ring of the ThP2. The structure will form a basis for future mechanistic and structural studies, as well as engineering aimed at production of alternative beta-amino acids.

PubMed: 14623876
DOI: 10.1074/JBC.M310803200

実験手法

X-RAY DIFFRACTION (2.45 Å)