The content of carboxyl-terminal fragments of parathormone in extracts of fresh bovine parathyroids.
Fresh parathyroid gland homogenates and fractions thereof were analyzed for their content of PTH and carboxyl-terminal fragments of the hormone. The tissue proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then extracted from gel fractions for RIA. Native PTH and PTH-(37-84) were used as standards to mark the migration positions of these peptides in the gels. The RIA for carboxyl-terminal hormone fragments used PTH-(37-84) as radioiodinated tracer and responded equally on a molar basis to either PTH or PTH-(37-84), making possible quantitative evaluation of both peptides in one assay after their separation. The results indicated that tissue homogenates contain 0.3-0.5 PTH-(37-84) moleq for each mole of PTH. Particulate fractions of the homogenates contained 0.15-0.3 moleq of fragment/mol PTH, while the high speed supernatant fraction of the homogenate contained about 2 moleq of fragment/mol PTH. When the experiments were performed using homogenization and fractionation buffers that contained numerous protease inhibitors, the ratios of carboxyl-terminal PTH fragment to intact hormone were not decreased, indicating that the hormone fragments were not produced during tissue processing. In addition, PTH added to tissue homogenates was not degraded during subsequent manipulations. The results demonstrate that fresh bovine parathyroid tissue contains substantial levels of carboxyl-terminal PTH peptide fragments, which can be measured by RIA after separation from PTH and other hormonal species. The data support the hypothesis that hormone fragments reside in regions of the cell different from those that contain PTH.
D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D010280:Parathyroid Glands / Q000032:analysis*; D010281:Parathyroid Hormone / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis*; D011863:Radioimmunoassay; D013347:Subcellular Fractions / Q000032:analysis
D010281:Parathyroid Hormone; D010446:Peptide Fragments; C037248:parathyroid hormone (37-84)