title: Pentavalent ions dependency of mammalian adenosine kinase.
abstract: The enzyme adenosine kinase (AK) has been purified to homogeneity from Syrian hamster and bovine livers. The purified enzymes from both these sources have a Mr of approximately 38 kDa, as determined by gel-filtration and SDS-polyacrylamide gel electrophoresis. A novel characteristic of AK observed here is that its catalytic activity shows a nearly complete dependence upon the presence of pentavalent ions such as phosphate (P(i)), arsenate or vanadate. Maximal AK activity was observed in the presence of either 2-3 mM P(i), or 5-10 mM arsenate, or 10-20 mM vanadate. A low basal level of AK activity (1-5% of maximal) observed in the absence of these ions is attributed to P(i) contamination in the adenine nucleotides preparations. The presence of P(i) had no effect on the K m for ATP (0.4 mM), but it markedly increased the affinity of the enzyme for adenosine. The K(m) of AK for adenosine in presence of 0, 0.1 mM and 2 mM P(i) was estimated to be 1.4 mu M, 0.77 mu M and 0.095 mu M, respectively. Free P(i) showed no exchange with any of the reactants during the assay conditions, and its presence had no effect on the thermostability of the enzyme. These observations suggest that the pentavalent ions such as phosphate may be playing an important role in the enzyme's catalytic mechanism by facilitating either binding of adenosine to the enzyme or in the formation of an enzyme-ATP-adenosine complex.
authors.affiliation: Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
authors.forename: W
authors.initials: W
authors.lastname: Hao
authors.forename: R S
authors.initials: RS
authors.lastname: Gupta
mesh_terms: D000248:Adenosine Kinase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001149:Arsenates; D002417:Cattle; D006224:Cricetinae; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008647:Mesocricetus; D014638:Vanadates
chemical_list: D001149:Arsenates; D014638:Vanadates; D000248:Adenosine Kinase; C025657:arsenic acid