title: Kinetic and hysteretic behavior of ATP hydrolysis of the highly stable dimeric ATP synthase of Polytomella sp.
abstract: The F1FO-ATP synthase of the colorless alga Polytomella sp. exhibits a robust peripheral arm constituted by nine atypical subunits only present in chlorophycean algae. The isolated dimeric enzyme exhibits a latent ATP hydrolytic activity which can be activated by some detergents. To date, the kinetic behavior of the algal ATPase has not been studied. Here we show that while the soluble F1 sector exhibits Michaelis-Menten kinetics, the dimer exhibits a more complex behavior. The kinetic parameters (Vmax and Km) were obtained for both the F1 sector and the dimeric enzyme as isolated or activated by detergent, and this activation was also seen on the enzyme reconstituted in liposomes. Unlike other ATP synthases, the algal dimer hydrolyzes ATP on a wide range of pH and temperature. The enzyme was inhibited by oligomycin, DCCD and Mg-ADP, although oligomycin induced a peculiar inhibition pattern that can be attributed to structural differences in the algal subunit-c. The hydrolytic activity was temperature-dependent and exhibited activation energy of 4 kcal/mol. The enzyme also exhibited a hysteretic behavior with a lag phase strongly dependent on temperature but not on pH, that may be related to a possible regulatory role in vivo.
authors.affiliation: Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México, D.F., Mexico.
authors.forename: Alexa
authors.initials: A
authors.lastname: Villavicencio-Queijeiro
authors.affiliation: Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, México, D.F., Mexico.
authors.forename: Juan Pablo
authors.initials: JP
authors.lastname: Pardo
authors.affiliation: Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México, D.F., Mexico. Electronic address: dhalphen@ifc.unam.mx.
authors.forename: Diego
authors.initials: D
authors.lastname: González-Halphen
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism*; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D019281:Dimerization; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009840:Oligomycins / Q000494:pharmacology; D059748:Proteolysis; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016823:Volvocida / Q000201:enzymology*
chemical_list: D004791:Enzyme Inhibitors; D009840:Oligomycins; D004024:Dicyclohexylcarbodiimide; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases
keywords: Chlorophycean algae; Dimeric complex V; Hysteresis; Mitochondrial F(1)F(O)-ATP synthase; Polytomella sp.