Calcium-regulated biosynthesis of the parathyroid secretory protein, proparathyroid hormone, and parathyroid hormone in dispersed bovine parathyroid cells.
Dispersed bovine parathyroid cells were incubated in vitro with 3H-labeled amino acids for 30--120 min. Analysis of cell extracts by sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis revealed a calcium-regulated incorporation of 3H-labeled amino acids into three major peaks, one with a molecular weight (Mr) of 70,000 and two peaks in the 10,000 Mr region, and a minor 5,000 Mr peak. Furthermore, [3H]mannose was incorporated into the 70,000 Mr peak, which corresponds to the parathyroid secretory protein (PSP) recognized as a glycoprotein. The two peaks in the 10,000 Mr region had the electrophoretic mobility of [3H]bovine proparathyroid hormone (ProPTH) and [3H]bovine PTH (1--84), respectively. Immunoreactive PTH was only detected in a peak comigrating with [3H]PTH-(1--84). The nonimmunoreactive 5,000 Mr peptide was not further identified. The incorporation of radioactive amino acids into PSP, ProPTH, PTH-(1--84), and the 5000-daltons peptide was inversely related to the calcium concentration in the incubation medium. The incorporation of radioactive mannose into PSP was also greater at low extracellular calcium concentrations. Our findings provide evidence for an inverse relationship between the extracellular calcium concentration and the formation of PSP, ProPTH, PTH-(1--84), and a yet to be identified 5000-dalton peptide in dispersed bovine parathyroid cells.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000502:physiology*; D002135:Calcium-Binding Proteins / Q000096:biosynthesis*; D002417:Cattle; D053379:Chromogranin A; D002864:Chromogranins; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D010280:Parathyroid Glands / Q000378:metabolism*; D010281:Parathyroid Hormone / Q000096:biosynthesis*; D011498:Protein Precursors / Q000096:biosynthesis*
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D053379:Chromogranin A; D002864:Chromogranins; D010281:Parathyroid Hormone; D011498:Protein Precursors; C003219:proparathormone; D002118:Calcium