title: Role of the divalent metal ion in the NAD:malic enzyme reaction: an ESEEM determination of the ground state conformation of malate in the E:Mn:malate complex.
abstract: The conformation of L-malate bound at the active site of Ascaris suum malic enzyme has been investigated by electron spin echo envelope modulation spectroscopy. Dipolar interactions between Mn2+ bound to the enzyme active site and deuterium specifically placed at the 2-position, the 3R-position, and the 3S-position of L-malate were observed. The intensities of these interactions are related to the distance between each deuterium and Mn2+. Several models of possible Mn-malate complexes were constructed using molecular graphics techniques, and conformational searches were conducted to identify conformers of malate that meet the distance criteria defined by the spectroscopic measurements. These searches suggest that L-malate binds to the enzyme active site in the trans conformation, which would be expected to be the most stable conformer in solution, not in the gauche conformer, which would be more similar to the conformation required for oxidative decarboxylation of oxalacetate formed from L-malate at the active site of the enzyme.
authors.affiliation: Department of Biochemistry, University of Missouri-Columbia 65211, USA. muccgw.bctipton@ssgate.missouri.edu
authors.forename: P A
authors.initials: PA
authors.lastname: Tipton
authors.forename: T P
authors.initials: TP
authors.lastname: Quinn
authors.forename: J
authors.initials: J
authors.lastname: Peisach
authors.forename: P F
authors.initials: PF
authors.lastname: Cook
mesh_terms: D000818:Animals; D017165:Ascaris suum; D001665:Binding Sites; D002413:Cations, Divalent / Q000737:chemistry; D003903:Deuterium; D004578:Electron Spin Resonance Spectroscopy; D008291:Malate Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D008293:Malates / Q000737:chemistry*; D008345:Manganese / Q000737:chemistry* / Q000378:metabolism; D009243:NAD / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D013113:Spin Labels / Q000138:chemical synthesis; D013237:Stereoisomerism; D013329:Structure-Activity Relationship
chemical_list: D002413:Cations, Divalent; D008293:Malates; D013113:Spin Labels; D009243:NAD; D008345:Manganese; C030298:malic acid; D003903:Deuterium; D008291:Malate Dehydrogenase; C046446:malate dehydrogenase-(oxaloacetate-decarboxylating) (NAD+)