![]() ![]() The ATP concentration ranged from 0 to 3 mM. The assay mixture contains 40 mM malate, 10 mM MgCl 2, and 1 mM NAD + or NADP +. The inhibited enzyme activities were assayed with NAD + ( A) or NADP + ( B) as the cofactor. Inhibitory effect of ATP on human WT and mutant c-NADP-ME. Kinetic parameters for human wild-type and nucleotide-binding mutant variants of c-NADP-ME. These figures were generated using PyMOL (DeLano Scientific LLC, San Carlos, CA). The yellow dashed lines represent the polar contacts between amino acid residues and NADP +. Ser346, Lys347 and Lys362 are colored green, purple and pink, respectively, and they are shown in a ball-and-stick model. ( C) The binding mode of NADP + in the active site of c-NADP-ME (PDB code 1GQ2). This figure was generated using the BioEdit sequence alignment editor program. Amino acid sequences of malic enzymes were searched using BLAST, and the alignments were generated by Clustal W. ( B) Multiple sequence alignments of three clusters of malic enzyme isoforms around the nucleotide-binding region of the active site. NADP + in the active site is colored yellow, and the Mn 2+ ion is red they are displayed in the sphere model. The active site with Mn 2+ and NADP + in each subunit is indicated. ( A) Tetramer of pigeon c-NADP-ME (PDB code 1GQ2). ![]()
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