Prolidase in human breast cancer MCF-7 cells.
Prolidase (EC 3.4.13.9) is an ubiquitously distributed imidodipeptidase that catalyzes the hydrolysis of dipeptides containing C-terminal proline or hydroxyproline. The enzyme plays an important role in the recycling of proline for collagen synthesis and cell growth. We have shown previously that prolidase activity in normal human skin fibroblasts is regulated by the interaction of type I collagen with beta1 integrin receptor. In the present study, we investigate prolidase activity in MCF-7 cells and find it is only one-third of that in normal human skin fibroblasts. The relative difference in prolidase activity is corroborated by enzyme protein with Western immunoblot analysis. We propose that the decrease in prolidase activity is due to derangement of regulation by the collagen-beta1 integrin receptor axis. Supporting evidence comes from the following observations: (1) relative collagen content elaborated by MCF-7 cells as compared to fibroblasts is lower by 30% in sparse cells and by 80% at confluence; (2) collagenase treatment of both cell types results in decreased enzyme activity; (3) in contrast to fibroblasts, prolidase activity in MCF-7 cells is not stimulated by the addition of type I collagen or beta1 integrin antibodies (agonist for beta1 integrin receptor); and (4) in contrast to fibroblasts, MCF-7 cells express only trace amounts of beta1 integrin receptor as shown by Western immunoblot analysis. Thus, we conclude that depressed prolidase activity in MCF-7 cells may be a result of disturbances in signaling mediated by beta1 integrin-collagen interaction.
Palka
J A
JA
Laboratory of Nutritional and Molecular Regulation, Division of Basic Sciences, National Cancer Institute-Frederick Cancer Research and Development Center, MD 21702, USA.
Phang
J M
JM
D000906:Antibodies / Q000494:pharmacology; D001943:Breast Neoplasms / Q000201:enzymology*; D002478:Cells, Cultured; D002648:Child; D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000494:pharmacology; D004150:Dipeptidases / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D019012:Integrin beta1 / Q000276:immunology; D008297:Male; D014407:Tumor Cells, Cultured
D000906:Antibodies; D019012:Integrin beta1; D003094:Collagen; D004150:Dipeptidases; C022449:proline dipeptidase; D017364:Collagenases