1N0H

Crystal Structure of Yeast Acetohydroxyacid Synthase in Complex with a Sulfonylurea Herbicide, Chlorimuron Ethyl

Summary for 1N0H

• Entry DOI: 10.2210/pdb1n0h/pdb
• Related: 1JSC
• Descriptor: Acetolactate synthase, POTASSIUM ION, MAGNESIUM ION, ... (9 entities in total)
• Functional Keywords: acetohydroxyacid synthase, sulfonylurea, herbicide inhibition, thiamine diphosphate, lyase
• Biological source: Saccharomyces cerevisiae (baker's yeast)
• Cellular location: Mitochondrion: P07342
• Total number of polymer chains: 2
• Total formula weight: 150838.92

Authors

Pang, S.S.,Guddat, L.W.,Duggleby, R.G. (deposition date: 2002-10-14, release date: 2003-01-07, Last modification date: 2024-11-06)

Primary citation

Pang, S.S.,Guddat, L.W.,Duggleby, R.G.
Molecular basis of sulfonylurea herbicide inhibition of acetohydroxyacid synthase
J.BIOL.CHEM., 278:7639-7644, 2003

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: 14557277
DOI: 10.1074/jbc.M211648200

Experimental method

X-RAY DIFFRACTION (2.8 Å)