Stimulation of guanylate cyclase in rat parotid membranes by phosphate.
The natriuretic peptide receptor guanylate cyclases are members of the membrane-bound guanylate cyclase family. Atrial natriuretic pepticle (ANP) stimulates guanylate cyclase A, which is predominantly expressed in the rat parotid gland. ATP is well known to increase ANP-stimulated guanylate cyclase, with Mg2+ as a cofactor. We investigated the regulation of guanylate cyclase activity in rat parotid membranes with Mn2+ as a cofactor, because enzyme activity was much higher with Mn2+ than with Mg2+. ANP (10(-7)M) stimulated guanylate cyclase activity to 120%-130% of the control level, and ATP (0.1-1.0 mM) depressed its activity, with or without ANP, to 70%-80% of the control. Enzyme activity was increased by the addition of phosphate (5-20 mM). In the presence of phosphate (5 mM), guanylate cyclase with and without ANP was maximally stimulated to 5- and 6.6-fold of the control, respectively. The net stimulation of guanylate cyclase by ANP was increased at ATP concentrations between 0.2 and 0.5 mM in the presence of phosphate (5 mM or 10 mM), but no change was observed in the absence of phosphate. Phosphate not only stimulated guanylate cyclase in the absence of ATP but altered the ATP regulation of ANP-stimulated guanylate cyclase. The stimulation of guanylate cyclase by phosphate may depend on a peculiarity of rat parotid membranes.
Nashida
Tomoko
T
Department of Biochemistry, The Nippon Dental University School of Dentistry at Niigata, 1-8 Hamaura-cho, Niigata, Japan. nashida@ngt.ndu.ac.jp.
Imai
Akane
A
Shimomura
Hiromi
H
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000494:pharmacology; D006152:Cyclic GMP / Q000032:analysis; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors / Q000187:drug effects*; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D008345:Manganese / Q000494:pharmacology; D008565:Membrane Proteins / Q000187:drug effects*; D010306:Parotid Gland / Q000187:drug effects / Q000201:enzymology*; D010710:Phosphates / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar
D008565:Membrane Proteins; D010710:Phosphates; D008345:Manganese; D009320:Atrial Natriuretic Factor; D000255:Adenosine Triphosphate; D006162:Guanylate Cyclase; D006152:Cyclic GMP; D008274:Magnesium