Procalcitonin is a glycoprotein
Journal of Biological Chemistry
Messenger RNA extracted from rat medullary carcinoma of the thyroid directs the synthesis in cell-free translation systems of a precursor of calcitonin, Mr = 15,000, substantially larger than the mature form of the hormone, Mr = 3,500. When translations of the mRNA were carried out in the presence of microsomal membranes prepared from a canine pancreas, a larger product (apparent Mr = 17,000) was observed by electrophoresis of the labeled proteins in the translation mixtures on sodium dodecyl
... on sodium dodecyl sulfate-polyacrylamide gels. This membrane-processed product of Mr = 17,000 was specifically immunoprecipitated by an antiserum to synthetic calcitonin and bound to concanavalin A-Sepharose. Incubation of the proteins synthesized in the cell-free translations performed in the presence of microsomal membranes with the glycosidase, endo-beta-N-acetylglucosaminidase H, reduced the apparent molecular weight of the membrane-processed precursor from 17,000 to 12,000. In addition, the processed Mr = 17,000 calcitonin-related precursor, but not the initial, unprocessed precursor of Mr = 15,000, was resistant to proteolytic digestion by a mixture of trypsin and chymotrypsin. These results indicate that the biosynthesis of calcitonin involves the glycosylation and proteolytic cleavage of a newly synthesized precursor along with sequestration of the processed precursor within microsomal vesicles. Thus, the calcitonin precursor undergoes extensive co- and post-translational processing to the smaller, unglycosylated hormone that is secreted.