Crystal structure of NAD-dependent malate dehydrogenase complexed with NADP(H) |
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Authors: | Tomita Takeo Fushinobu Shinya Kuzuyama Tomohisa Nishiyama Makoto |
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Affiliation: | Biotechnology Research Center, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan. |
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Abstract: | For better understanding of the coenzyme specificity in NAD-dependent MDH (tMDH) from Thermus flavus AT-62, we determined the crystal structures of tMDH-NADP(H) complex at maximally 1.65 A resolution. The overall structure is almost the same as that of the tMDH-NADH complex. However, NADP(H) binds to tMDH in the reverse orientation, where adenine occupies the position near the catalytic center and nicotinamide is positioned at the adenine binding site of the tMDH-NADH complex. Consistent with this, kinetic analysis of the malate-oxidizing reaction revealed that NADP(+) inhibited tMDH at high concentrations. This has provided the first evidence for the alternative binding mode of the nicotinamide coenzyme, that has pseudo-symmetry in its structure, in a single enzyme. |
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Keywords: | Crystal structure Malate dehydrogenase Coenzyme specificity Tricarboxylic acid cycle |
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