2HDB
HMG-CoA synthase from Enterococcus faecalis. Mutation alanine 110 to glycine
Summary for 2HDB
| Entry DOI | 10.2210/pdb2hdb/pdb |
| Related | 1TVZ 1TXT 1X9E 1XPK 1XPM 1YSL |
| Descriptor | HMG-CoA synthase, SULFATE ION, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, ... (4 entities in total) |
| Functional Keywords | thiolase fold, synthase, protein, mutant, lyase |
| Biological source | Enterococcus faecalis |
| Total number of polymer chains | 2 |
| Total formula weight | 85033.75 |
| Authors | Steussy, C.N.,Sutherlin, A.,Stauffacher, C.V. (deposition date: 2006-06-20, release date: 2007-03-20, Last modification date: 2023-08-30) |
| Primary citation | Steussy, C.N.,Robison, A.D.,Tetrick, A.M.,Knight, J.T.,Rodwell, V.W.,Stauffacher, C.V.,Sutherlin, A.L. A structural limitation on enzyme activity: the case of HMG-CoA synthase. Biochemistry, 45:14407-14414, 2006 Cited by PubMed Abstract: Recent structural studies of the HMG-CoA synthase members of the thiolase superfamily have shown that the catalytic loop containing the nucleophilic cysteine follows the phi and psi angle pattern of a II' beta turn. However, the i + 1 residue is conserved as an alanine, which is quite unusual in this position as it must adopt a strained positive phi angle to accommodate the geometry of the turn. To assess the effect of the conserved strain in the catalytic loop, alanine 110 of Enterococcus faecalis 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase was mutated to a glycine. Subsequent enzymatic studies showed that the overall reaction rate of the enzyme was increased 140-fold. An X-ray crystallographic study of the Ala110Gly mutant enzyme demonstrated unanticipated adjustments in the active site that resulted in additional stabilization of all three steps of the reaction pathway. The rates of acetylation and hydrolysis of the mutant enzyme increased because the amide nitrogen of Ser308 shifts 0.4 A toward the catalytic cysteine residue. This motion positions the nitrogen to better stabilize the intermediate negative charge that develops on the carbonyl oxygen of the acetyl group during both the formation of the acyl-enzyme intermediate and its hydrolysis. In addition, the hydroxyl of Ser308 rotates 120 degrees to a position where it is able to stabilize the carbanion intermediate formed by the methyl group of the acetyl-S-enzyme during its condensation with acetoacetyl-CoA. PubMed: 17128980DOI: 10.1021/bi061505q PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.2 Å) |
Structure validation
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