title: Thermodynamics of binding of divalent magnesium and manganese to uridine phosphates: implications for diabetes-related hypomagnesaemia and carbohydrate biocatalysis.
abstract: BACKGROUND
  Although the necessity of divalent magnesium and manganese for full activity of sugar nucleotidyltransferases and glycosyltransferases is well known, the role of these metal cations in binding the substrates (uridine 5'-triphosphate, glucose-1-phosphate, N-acetylglucosamine-1-phosphate, and uridine 5'-diphosphate glucose), products (uridine 5'-diphosphate glucose, uridine 5'-diphosphate N-acetylglucosamine, pyrophosphate, and uridine 5'-diphosphate), and/or enzyme is not clearly understood.
  
  
  RESULTS
  Using isothermal titration calorimetry we have studied the binding relationship between the divalent metals, magnesium and manganese, and uridine 5'-phosphates to determine the role these metals play in carbohydrate biosynthesis. It was determined from the isothermal titration calorimetry (ITC) data that Mg(+2) and Mn(+2) are most tightly bound to PP(i), K(b) = 41,000 +/- 2000 M(-1) and 28,000 +/- 50,000 M(-1) respectively, and UTP, K(b) = 14,300 +/- 700 M(-1) and 13,000 +/- 2,000 M(-1) respectively.
  
  
  CONCLUSIONS
  Our results indicate that the formal charge state of the phosphate containing substrates determine the binding strength. Divalent metal cations magnesium and manganese showed similar trends in binding to the sugar substrates. Enthalpy of binding values were all determined to be endothermic except for the PP(i) case. In addition, entropy of binding values were all found to be positive. From this data, we discuss the role of magnesium and manganese in both sugar nucleotidyltransferase and glycosyltransferase reactions, the differences in metal-bound substrates expected under normal physiological metal concentrations and those of hypomagnesaemia, and the implications for drug design.
authors.affiliation: Department of Chemistry, Iowa State University, Ames, IA 50011-3111, USA. czea@gvc.edu
authors.forename: Corbin J
authors.initials: CJ
authors.lastname: Zea
authors.forename: Gulden
authors.initials: G
authors.lastname: Camci-Unal
authors.forename: Nicola L
authors.initials: NL
authors.lastname: Pohl