Raynaud F, Bauvois B, Gerbaud P, Evain-Brion D
Laboratoire de Physiopathologie du Développement, CNRS, URA 1337, Ecole Normale Supérieure, Paris, France.
J Cell Physiol. 1992 May;151(2):378-85. doi: 10.1002/jcp.1041510219.
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.
对人皮肤成纤维细胞进行细胞表面蛋白酶活性检测。一种从甘氨酰 - 脯氨酰 - 对硝基苯胺（Gly - Pro - pNA）的NH2末端去除二肽的活性，被二异丙基氟磷酸酯（DFP）、苯甲基磺酰氟（PMSF）和二丙基丁二酰胺（diprotin A）特异性抑制，因此被鉴定为二肽基肽酶IV（DPP IV）。当以丙氨酰 - 、亮氨酰 - 和精氨酰 - 对硝基苯胺（Ala - 、Leu - 、Arg - pNA）作为显色底物时，还对一组贝司他汀敏感的N - 外切氨基肽酶活性进行了表征。使用分别识别N - 丙氨酰氨基肽酶和DPP IV的抗CD 13和抗CD 26人单克隆抗体，发现人皮肤成纤维细胞在其表面表达CD 13和CD 26抗原。此外，两种肽酶都被抗CD 13和抗CD 26单克隆抗体从质膜上特异性免疫沉淀。还对来自皮肤病和风湿性疾病（即银屑病、类风湿性关节炎和扁平苔藓）的人成纤维细胞的细胞表面蛋白水解活性进行了研究。发现这些成纤维细胞也表达最初在正常皮肤成纤维细胞上鉴定出的两种蛋白酶类型，并且在所有情况下丙氨酰氨基肽酶活性水平相似。相比之下，三种病理性成纤维细胞群体中的精氨酰 - 、亮氨酰 - 外切氨基肽酶和DPP IV活性水平比其正常对应物显著更高（高达6.6倍）。因此，这些蛋白水解酶有可能作为皮肤病的标志物。综上所述，我们的结果表明，与丝氨酸和N - 氨基肽酶活性相关的皮肤成纤维细胞衍生的蛋白酶可能通过参与与成纤维细胞行为相关的细胞外事件发挥重要作用。
