1OZH
The crystal structure of Klebsiella pneumoniae acetolactate synthase with enzyme-bound cofactor and with an unusual intermediate.
Summary for 1OZH
| Entry DOI | 10.2210/pdb1ozh/pdb |
| Related | 1OZF 1OZG |
| Descriptor | Acetolactate synthase, catabolic, PHOSPHATE ION, MAGNESIUM ION, ... (7 entities in total) |
| Functional Keywords | acetolactate synthase, acetohydroxyacid synthase, thiamin diphosphate, lyase |
| Biological source | Klebsiella pneumoniae |
| Total number of polymer chains | 4 |
| Total formula weight | 248604.55 |
| Authors | Pang, S.S.,Duggleby, R.G.,Schowen, R.L.,Guddat, L.W. (deposition date: 2003-04-09, release date: 2003-11-04, Last modification date: 2023-08-16) |
| Primary citation | Pang, S.S.,Duggleby, R.G.,Schowen, R.L.,Guddat, L.W. The Crystal Structures of Klebsiella pneumoniae Acetolactate Synthase with Enzyme-bound Cofactor and with an Unusual Intermediate. J.Biol.Chem., 279:2242-2253, 2004 Cited by PubMed Abstract: Acetohydroxyacid synthase (AHAS) and acetolactate synthase (ALS) are thiamine diphosphate (ThDP)-dependent enzymes that catalyze the decarboxylation of pyruvate to give a cofactor-bound hydroxyethyl group, which is transferred to a second molecule of pyruvate to give 2-acetolactate. AHAS is found in plants, fungi, and bacteria, is involved in the biosynthesis of the branched-chain amino acids, and contains non-catalytic FAD. ALS is found only in some bacteria, is a catabolic enzyme required for the butanediol fermentation, and does not contain FAD. Here we report the 2.3-A crystal structure of Klebsiella pneumoniae ALS. The overall structure is similar to AHAS except for a groove that accommodates FAD in AHAS, which is filled with amino acid side chains in ALS. The ThDP cofactor has an unusual conformation that is unprecedented among the 26 known three-dimensional structures of nine ThDP-dependent enzymes, including AHAS. This conformation suggests a novel mechanism for ALS. A second structure, at 2.0 A, is described in which the enzyme is trapped halfway through the catalytic cycle so that it contains the hydroxyethyl intermediate bound to ThDP. The cofactor has a tricyclic structure that has not been observed previously in any ThDP-dependent enzyme, although similar structures are well known for free thiamine. This structure is consistent with our proposed mechanism and probably results from an intramolecular proton transfer within a tricyclic carbanion that is the true reaction intermediate. Modeling of the second molecule of pyruvate into the active site of the enzyme with the bound intermediate is consistent with the stereochemistry and specificity of ALS. PubMed: 14557277DOI: 10.1074/jbc.M304038200 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2 Å) |
Structure validation
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