1UPA
Carboxyethylarginine synthase from Streptomyces clavuligerus (SeMet structure)
Summary for 1UPA
| Entry DOI | 10.2210/pdb1upa/pdb |
| Related | 1UPB 1UPC |
| Descriptor | CARBOXYETHYLARGININE SYNTHASE, THIAMINE DIPHOSPHATE, MAGNESIUM ION, ... (5 entities in total) |
| Functional Keywords | synthase, clavulanic acid, antibiotic, lactamase |
| Biological source | STREPTOMYCES CLAVULIGERUS |
| Total number of polymer chains | 4 |
| Total formula weight | 248489.67 |
| Authors | Caines, M.E.C.,Elkins, J.M.,Hewitson, K.S.,Schofield, C.J. (deposition date: 2003-09-29, release date: 2003-11-20, Last modification date: 2024-10-16) |
| Primary citation | 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 Cited by 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: 14623876DOI: 10.1074/JBC.M310803200 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.35 Å) |
Structure validation
Download full validation report
