title: Direct Mg(2+) binding activates adenylate kinase from Escherichia coli.
abstract: We report evidence that adenylate kinase (AK) from Escherichia coli can be activated by the direct binding of a magnesium ion to the enzyme, in addition to ATP-complexed Mg(2+). By systematically varying the concentrations of AMP, ATP, and magnesium in kinetic experiments, we found that the apparent substrate inhibition of AK, formerly attributed to AMP, was suppressed at low magnesium concentrations and enhanced at high magnesium concentrations. This previously unreported magnesium dependence can be accounted for by a modified random bi-bi model in which Mg(2+) can bind to AK directly prior to AMP binding. A new kinetic model is proposed to replace the conventional random bi-bi mechanism with substrate inhibition and is able to describe the kinetic data over a physiologically relevant range of magnesium concentrations. According to this model, the magnesium-activated AK exhibits a 23- +/- 3-fold increase in its forward reaction rate compared with the unactivated form. The findings imply that Mg(2+) could be an important affecter in the energy signaling network in cells.
authors.affiliation: Department of Chemistry, University of California at Berkeley, Berkeley, California 94720.
authors.forename: Yan-Wen
authors.initials: YW
authors.lastname: Tan
authors.affiliation: Department of Chemistry, University of California at Berkeley, Berkeley, California 94720.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Hanson
authors.affiliation: Department of Chemistry, University of California at Berkeley, Berkeley, California 94720; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720. Electronic address: hawyang@berkeley.edu.
authors.forename: Haw
authors.initials: H
authors.lastname: Yang
mesh_terms: D000227:Adenine Nucleotides / Q000378:metabolism; D000263:Adenylate Kinase / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D005453:Fluorescence; D008024:Ligands; D008274:Magnesium / Q000378:metabolism*; D008958:Models, Molecular
chemical_list: D000227:Adenine Nucleotides; D008024:Ligands; D000263:Adenylate Kinase; D008274:Magnesium