Osteosarcoma cytosol factor promotes parathyroid hormone stimulation of adenylate cyclase independent of GTP.
The adenylate cyclase (ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1)-stimulating factor from rat osteosarcoma cytosol was purified 600-fold by ion-exchange chromatography. The factor has an apparent Mr of 20000, is cold-labile, but retains activity at -20 degrees C in 10% glycerol. The factor enhanced parathyroid hormone stimulation of adenylate cyclase and restored hormone responsiveness to membranes washed with 0.5 M NaCl. These 'GTP-like' effects were not inhibited by 100 microM GDP-beta-S, which completely abolished the GTP enhancement of both basal and hormone-stimulated adenylate cyclase. Adenylate cyclase activity in the presence of the stimulating factor was linear with time, and showed hyperbolic dependence on factor concentration. The factor also linearized (in double reciprocal plots) the downward-concave Mg2+-dependence of adenylate cyclase, increasing the apparent affinity of the enzyme for Mg2+. The presence of the factor in two clonal osteosarcoma cell lines correlated with parathyroid hormone-stimulatable adenylate cyclase. Factor stimulation was absent while GTP stimulation was retained in the hormone-nonresponsive clone. Factor and hormone sensitivity were restored by in vivo passage. This factor thus may represent a guanyl nucleotide-independent path for cellular regulation of hormone response.
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001688:Biological Products / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000502:physiology; D006160:Guanosine Triphosphate / Q000494:pharmacology*; D007700:Kinetics; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000503:physiopathology; D012516:Osteosarcoma / Q000503:physiopathology*; D010281:Parathyroid Hormone / Q000494:pharmacology*; D051381:Rats
D001688:Biological Products; D010281:Parathyroid Hormone; D006160:Guanosine Triphosphate; D000262:Adenylyl Cyclases