Characterization of specific proteases associated with the surface of human skin fibroblasts, and their modulation in pathology.
Human skin fibroblasts were probed for cell surface protease activity. One activity removing dipeptides from the NH2-terminal end of Gly-Pro-pNA was specifically inhibited by di-isopropyl-fluorophosphate (DFP), phenylmethanesulphony fluoride (PMSF), and diprotin A, and thus was identified as dipeptidyl peptidase IV (DPP IV). A group of bestatin-sensitive N-exoaminopeptidase activities was also characterized when Ala-, Leu-, and Arg-pNA were used as chromogenic substrates. Using human monoclonal antibodies anti-CD 13 and anti-CD 26 that recognized, respectively, an N-Ala-aminopeptidase and DPP IV, it was found that human dermal fibroblasts expressed the CD 13 and CD 26 antigen on their surface. In addition, both peptidases were specifically immunoprecipitated by monoclonal antibodies anti-CD 13 and anti-CD 26 from plasma membranes. Cell surface proteolytic activities were also investigated in human fibroblasts derived from dermatological and rheumatic diseases (i.e., psoriasis, rheumatoid arthritis, and lichen planus). It was found that these fibroblasts also expressed both types of proteinases initially identified on normal skin fibroblasts and that the levels of Ala-aminopeptidase activities were similar in all cases. In contrast, the levels of Arg-, Leu-exoaminopeptidase, and DPP IV activities were significantly higher (up to 6.6-fold) in the three pathological fibroblast populations than in their normal counterparts. These proteolytic enzymes, therefore, can potentially serve as markers in dermatological diseases. Taken together, our results suggest that skin fibroblast-derived proteinases associated with both serine and N-aminopeptidase activities may play an important role by participating in the extracellular events associated with fibroblast behaviour.
Raynaud
F
F
Laboratoire de Physiopathologie du Développement, CNRS, URA 1337, Ecole Normale Supérieure, Paris, France.
Bauvois
B
B
Gerbaud
P
P
Evain-Brion
D
D
D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007154:Immune System Diseases / Q000201:enzymology*; D011233:Precipitin Tests; D011480:Protease Inhibitors / Q000494:pharmacology; D012867:Skin / Q000201:enzymology* / Q000473:pathology; D013379:Substrate Specificity
D000911:Antibodies, Monoclonal; D011480:Protease Inhibitors; D010450:Endopeptidases