title: Kinetic studies of the effects of K+, Na+ and Li+ on the catalytic activity of human erythrocyte pyridoxal kinase.
abstract: Kinetic studies were conducted to examine the effects of K+, Na+ and Li+ on human erythrocyte pyridoxal kinase (PK) activity. A dialyzed hemolysate served as the PK source. The substrates used were pyridoxal (PL) and ATP. Determination of the enzymatic activity was based on HPLC separation and fluorimetric detection of PL and pyridoxal 5'-phosphate as semicarbazone derivatives. In comparison to the poor activity of PK assayed without monovalent cation, all tested cations are activators. Among them, K+ is the most effective, improving both PK affinity for the substrates and maximal velocity. Na+ increases maximal velocity and PK affinity for ATP but decreases it for PL. Li+ is a poor activator which seems to modify the enzymatic mechanism from a random to an ordered sequential pattern with ATP bound before PL. Results suggest that K+ and Na+ bind to PK on the same site while Li+ binds on another site. This hypothesis and the mechanism of monovalent cation-PK interaction are compared to other well-known K(+)-activated enzymes.
authors.affiliation: Laboratoire de Pharmacologie et Toxicologie, Centre Hospitalier universitaire, Angers, France.
authors.forename: P
authors.initials: P
authors.lastname: Lainé-Cessac
authors.forename: P
authors.initials: P
authors.lastname: Allain
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002414:Cations, Monovalent / Q000737:chemistry; D004912:Erythrocytes / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008094:Lithium / Q000737:chemistry*; D009994:Osmolar Concentration; D011188:Potassium / Q000737:chemistry*; D011730:Pyridoxal / Q000378:metabolism; D011731:Pyridoxal Kinase / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D012964:Sodium / Q000737:chemistry*
chemical_list: D002414:Cations, Monovalent; D011730:Pyridoxal; D000255:Adenosine Triphosphate; D008094:Lithium; D012964:Sodium; D011731:Pyridoxal Kinase; D011188:Potassium