A serum factor promotes collagenase synthesis by an osteoblastic cell line.
Regulation of the synthesis of collagenase was investigated in the osteoblastic cell line, UMR 106-01. The cells were stained by the avidin-biotin-complex technique for the presence of the enzyme. By this method, it was possible to identify cells producing collagenase. Synthesis, but not secretion, was found to be constitutive in these cells with the enzyme located intracellularly in cytoplasmic vesicles and the Golgi apparatus. The amount of collagenase contained within UMR cells and the number of cells synthesizing the enzyme were proportional to the concentration of fetal bovine serum in the incubating medium. When serum was withdrawn from the osteosarcoma cells, the content of collagenase decreased with time and the enzyme became undetectable by 48 h of serum depletion. The decrease in collagenase content could be completely reversed by resupplying serum to the cells. The collagenase promoting activity of serum could not be eliminated by adsorption on activated charcoal but was retained by a dialysis membrane with a 12,000 mol wt cutoff. A range of bone-seeking hormones or agents known to affect collagenase secretion was added to the medium in an attempt to mimic the effect of serum on collagenase accumulation. None of these agonists, including parathyroid hormone, could reproduce the effect of serum on these cells, although parathyroid hormone could act as a collagenase secretagogue in the presence or absence of serum. It is concluded that fetal bovine serum contains a yet unidentified factor or factors greater than 12,000 mol wt responsible for the continued synthesis of collagenase by UMR 106-01 cells.
Puccinelli
J M
JM
Pediatric Research Institute, Cardinal Glennon Children's Hospital, St. Louis, Missouri.
Omura
T H
TH
Strege
D W
DW
Jeffrey
J J
JJ
Partridge
N C
NC
D000818:Animals; D001798:Blood Proteins / Q000032:analysis / Q000494:pharmacology*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D007150:Immunohistochemistry; D003012:Microbial Collagenase / Q000096:biosynthesis*; D008970:Molecular Weight; D010006:Osteoblasts / Q000166:cytology / Q000201:enzymology*; D010281:Parathyroid Hormone / Q000494:pharmacology; D051381:Rats; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000494:pharmacology
D001798:Blood Proteins; D010281:Parathyroid Hormone; D012710:Serum Albumin, Bovine; D003012:Microbial Collagenase