Prolidase activity in fibroblasts is regulated by interaction of extracellular matrix with cell surface integrin receptors.
Prolidase (EC 3.4.13.9) is a ubiquitously distributed imidodipeptidase that catalyzes the hydrolysis of C-terminal proline or hydroxyproline containing dipeptides. The enzyme plays an important role in the recycling of proline for collagen synthesis and cell growth. An increase in enzyme activity is correlated with increased rates of collagen turnover indicative of extracellular matrix (ECM) remodeling, but the mechanism linking prolidase activity and ECM is poorly understood. Thus, the effect of ECM-cell interaction on intracellular prolidase activity is of special interest. In cultured human skin fibroblasts, the interaction with ECM and, more specifically, type I collagen mediated by the beta 1 integrin receptor regulates cellular prolidase activity. Supporting evidence comes from the following observations: 1) in sparse cells with a low amount of ECM collagen or in confluent cells in which ECM collagen was removed by collagenase (but not by trypsin or elastase) treatment, prolidase activity was decreased; 2) this effect was reversed by the addition of type I collagen or beta 1 integrin antibody (agonist for beta 1 integrin receptor); 3) sparse cells (with typically low prolidase activity) showed increased prolidase activity when grown on plates coated with type I collagen or on type IV collagen and laminin, constituents of basement membrane; 4) the relative differences in prolidase activity due to collagenase treatment and subsequent recovery of the activity by beta 1 integrin antibody or type I collagen treatment were accompanied by parallel differences in the amount of the enzyme protein recovered from these cells, as shown by Western immunoblot analysis. Thus, we conclude that prolidase activity responded to ECM metabolism (tissue remodeling) through signals mediated by the integrin receptor.
Palka
J A
JA
Laboratory of Nutritional and Molecular Regulation, National Cancer Institute-Frederick Cancer Research and Development Center, Maryland 21702, USA.
Phang
J M
JM
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002452:Cell Count; D002460:Cell Line; D002648:Child; D003094:Collagen / Q000378:metabolism / Q000494:pharmacology; D004150:Dipeptidases / Q000378:metabolism*; D004549:Elastin / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology*; D006801:Humans; D019012:Integrin beta1 / Q000276:immunology / Q000502:physiology*; D007700:Kinetics; D008297:Male; D012867:Skin
D000911:Antibodies, Monoclonal; D019012:Integrin beta1; D003094:Collagen; D004549:Elastin; D004150:Dipeptidases; C022449:proline dipeptidase