4kqx

Mutant Slackia exigua KARI DDV in complex with NAD and an inhibitor

spinqualitylabelsall models 4kqx, resolution 1.80Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 4kqx is a 2 chain structure with sequence from Slackia exigua ATCC 700122. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.8Å Ligands: HIO, HIS, MG, NAD Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function ILVC_SLAES Involved in the biosynthesis of branched-chain amino acids (BCAA). Catalyzes an alkyl-migration followed by a ketol-acid reduction of (S)-2-acetolactate (S2AL) to yield (R)-2,3-dihydroxy-isovalerate. In the isomerase reaction, S2AL is rearranged via a Mg-dependent methyl migration to produce 3-hydroxy-3-methyl-2-ketobutyrate (HMKB). In the reductase reaction, this 2-ketoacid undergoes a metal-dependent reduction by NADPH to yield (R)-2,3-dihydroxy-isovalerate.[HAMAP-Rule:MF_00435] Publication Abstract from PubMed To date, efforts to switch the cofactor specificity of oxidoreductases from nicotinamide adenine dinucleotide phosphate (NADPH) to nicotinamide adenine dinucleotide (NADH) have been made on a case-by-case basis with varying degrees of success. Here we present a straightforward recipe for altering the cofactor specificity of a class of NADPH-dependent oxidoreductases, the ketol-acid reductoisomerases (KARIs). Combining previous results for an engineered NADH-dependent variant of Escherichia coli KARI with available KARI crystal structures and a comprehensive KARI-sequence alignment, we identified key cofactor specificity determinants and used this information to construct five KARIs with reversed cofactor preference. Additional directed evolution generated two enzymes having NADH-dependent catalytic efficiencies that are greater than the wild-type enzymes with NADPH. High-resolution structures of a wild-type/variant pair reveal the molecular basis of the cofactor switch. General approach to reversing ketol-acid reductoisomerase cofactor dependence from NADPH to NADH.,Brinkmann-Chen S, Flock T, Cahn JK, Snow CD, Brustad EM, McIntosh JA, Meinhold P, Zhang L, Arnold FH Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):10946-51. doi:, 10.1073/pnas.1306073110. Epub 2013 Jun 17. PMID:23776225 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See Also Ketol-acid reductoisomerase 3D structures

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