Comparison of the biochemical properties, regulation and function of ATP-diphosphohydrolase from human placenta and rat kidney.
ATP-diphosphohydrolase (apyrase. EC 3.6.1.5) has both ATPase and ADPase activity that are stimulated by bivalent metals, with Ca2+ being the most effective. The possible physiological function of this enzyme, associated with placental and renal microvilli, is related to the extracellular metabolism of nucleotides. A comparison of the biochemical properties of human placenta and rat kidney apyrase is presented, showing similarities in Mr. bivalent metal stimulation, nucleotide nonspecificity, insensitivity towards specific ATPase inhibitors, and lack of essential sulfhydryl and aliphatic hydroxyl groups. We describe the treatment of membrane preparations from both tissues with different detergents and the isoelectric focusing of the solubilized proteins to partially purify apyrase. An ectoenzyme localization is assigned both in microvillus membranes and in the vasculature on the basis of organ perfusion experiments with nucleotides in the presence of antibodies. Placental and kidney microvillus membranes inhibited ADP-induced platelet aggregation, in agreement with an extracellular role. Initial studies on enzyme regulation suggested the existence of at least two types of modulatory proteins: an activating protein in the cytosol of both tissues, and an inhibitory protein associated with placental microsomes. Possible hormonal regulation was investigated in kidneys using in vivo estradiol treatment, but only slight changes in total apyrase activity were observed.
Valenzuela
M A
MA
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chile.
Kettlun
A M
AM
Sandoval
S
S
García
L
L
Mancilla
M
M
Neckelmann
G
G
Chayet
L
L
Alvarez
A
A
Cuevas
F
F
Collados
L
L
Espinosa
V
V
Traverso-Cori
A
A
Bravo
I
I
Acevedo
C G
CG
Aranda
E
E
D000818:Animals; D001081:Apyrase / Q000737:chemistry / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology; D006801:Humans; D007668:Kidney / Q000201:enzymology*; D010920:Placenta / Q000201:enzymology*; D010974:Platelet Aggregation / Q000187:drug effects; D051381:Rats
D004958:Estradiol; D001081:Apyrase