The calcitonin receptor expressed by the porcine LLC-PK1 renal tubule cells is a seven-transmembrane domain, G protein-coupled receptor activating adenylyl-cyclase and phospholipase C. Salmon calcitonin stimulated dose- and time-dependent release of the phospholipase D-dependent phosphatidylcholine product +AFs-H-3+AF0-choline with an EC50 +AD0- 2.5 0.3 x 10(-8) M, similar to that determined for phosphoinositide metabolism (EC50 +AD0- 4.5 1.0 x 10(-8) M). The hormone failed to induce release of +AFs-H-3+AF0- phosphocholine and +AFs-H-3+AF0-glycerophosphocholine, ruling out activation of phosphatydilcholine-specific phospholipase C and phospholipase A. Calcitonin stimulated phosphatidic acid, a product of phospholipase D-dependent phosphatydilcholine hydrolysis. Activation of phospholipase D was confirmed by release of +AFs-H-3+AF0-phosphatydilethanol, a specific and stable product in the presence of a primary alcohol. Activation of calcitonin receptor induced diacylglycerol formation, with a rapid peak followed by a prolonged increase, due to activation of phospholipase C and of phospholipase D. Consequently, the protein kinase-C alpha, but not the delta isoenzyme, was cytosol-to-membrane translocated by approximately 50+ACU- after 20 min exposure to calcitonin, whereas protein kinase-C zeta, which was approximately 40+ACU- membrane-linked in unstimulated cells, translocated by approximately 19+ACU-. The human calcitonin receptor expressed by BIN-67 ovary tumor cells, although displaying higher affinity for calcitonin, failed to activate phospholipase D and protein kinase-C in response to the hormone. This receptor lacks the G protein binding consensus site due to the presence of a 48-bp cassette encoding for a 16-amino acid insert in the predicted first intracellular loop. This modification is likely to prevent the calcitonin receptor from associating to phospholipase-coupled signaling.

Phospholipase D- and protein kinase C isoenzyme-dependent signal transduction pathways activated by the calcitonin receptor

TETI, ANNA MARIA;
1998-01-01

Abstract

The calcitonin receptor expressed by the porcine LLC-PK1 renal tubule cells is a seven-transmembrane domain, G protein-coupled receptor activating adenylyl-cyclase and phospholipase C. Salmon calcitonin stimulated dose- and time-dependent release of the phospholipase D-dependent phosphatidylcholine product +AFs-H-3+AF0-choline with an EC50 +AD0- 2.5 0.3 x 10(-8) M, similar to that determined for phosphoinositide metabolism (EC50 +AD0- 4.5 1.0 x 10(-8) M). The hormone failed to induce release of +AFs-H-3+AF0- phosphocholine and +AFs-H-3+AF0-glycerophosphocholine, ruling out activation of phosphatydilcholine-specific phospholipase C and phospholipase A. Calcitonin stimulated phosphatidic acid, a product of phospholipase D-dependent phosphatydilcholine hydrolysis. Activation of phospholipase D was confirmed by release of +AFs-H-3+AF0-phosphatydilethanol, a specific and stable product in the presence of a primary alcohol. Activation of calcitonin receptor induced diacylglycerol formation, with a rapid peak followed by a prolonged increase, due to activation of phospholipase C and of phospholipase D. Consequently, the protein kinase-C alpha, but not the delta isoenzyme, was cytosol-to-membrane translocated by approximately 50+ACU- after 20 min exposure to calcitonin, whereas protein kinase-C zeta, which was approximately 40+ACU- membrane-linked in unstimulated cells, translocated by approximately 19+ACU-. The human calcitonin receptor expressed by BIN-67 ovary tumor cells, although displaying higher affinity for calcitonin, failed to activate phospholipase D and protein kinase-C in response to the hormone. This receptor lacks the G protein binding consensus site due to the presence of a 48-bp cassette encoding for a 16-amino acid insert in the predicted first intracellular loop. This modification is likely to prevent the calcitonin receptor from associating to phospholipase-coupled signaling.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11697/5977
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