Constitutive synthesis of a 92-kDa keratinocyte-derived type IV collagenase is enhanced by type I collagen and decreased by type IV collagen matrices.
Constitutive synthesis of a 92-kDa keratinocyte-derived type IV collagenase is enhanced by type I collagen and decreased by type IV collagen matrices.
复制标题
I 型胶原增强了 92 kDa 角质细胞衍生的 IV 型胶原酶的组成型合成，而 IV 型胶原基质则减少了这种合成。
DOI：
10.1111/1523-1747.ep12614800
复制
发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Wynn,KC
中科院分区：
文献类型：
--
作者：
Sarret,Y;Woodley,DT;Goldberg,GS;Kronberger,A;Wynn,KC
人角质形成细胞合成间质胶原酶、72-kDa 明胶酶和最近描述的 92-kDa 明胶酶/IV 型胶原酶。我们检查了与塑料、基底膜胶原蛋白（IV 型）和间质真皮胶原蛋白（I 型）相对应的基底角质细胞合成这种新型酶的情况。条件培养基样品在 10% 聚丙烯酰胺、明胶负载酶谱上进行电泳。通过凝胶澄清来鉴定具有明胶切割特性的蛋白质条带，并通过光密度测定法进行定量。 92 kDa 条带具有显着的明胶分解活性，并且在培养 72 小时后会增加。使用针对 92-kDa IV 型胶原酶的单特异性抗体通过蛋白质免疫印迹法证明了该 92-kDa 条带为 IV 型胶原酶。与使用 IV 型胶原的培养物相比，使用 I 型胶原的角质形成细胞的 92-kDa 酶合成量增加了三倍，与塑料相比增加了 1.5 倍。这种增强的特异性通过其他蛋白质（例如 72-kDa 明胶酶）的恒定水平得到体现。这项研究表明，细胞-基质相互作用调节最近描述的角质细胞衍生的 92-kDa 明胶酶的合成，并且特定的胶原蛋白类型（I 与 IV）对该酶的合成具有相反的影响。
Human keratinocytes synthesize interstitial collagenase, a 72-kDa gelatinase, and a recently described 92-kDa gelatinase/type IV collagenase. We examined the synthesis of this novel enzyme by basal keratinocytes apposed to plastic, basement membrane collagen (type IV), and interstitial dermal collagen (type I). Samples of conditioned medium were electrophoresed on a 10% polyacrylamide, gelatin-ladened zymogram. Protein bands with gelatin-cleaving properties were identified by clarification of the gel and quantified by densitometry. A 92-kDa band had marked gelatinolytic activity and increased in culture over 72 h. The identification of this 92-kDa band as type IV collagenase was demonstrated by Western immunoblotting using monospecific antibody to the 92-kDa type IV collagenase. Keratinocytes apposed to type I collagen exhibited a threefold increase in the synthesis of the 92-kDa enzyme compared to cultures apposed to type IV collagen and a 1.5-times increase compared to plastic. The specificity of this enhancement was shown by constant levels of other proteins (e.g., the 72-kDa gelatinase). This study demonstrates that cell-matrix interactions modulate the synthesis of a recently described, keratinocyte-derived, 92-kDa gelatinase and that specific collagen types (I versus IV) have opposite effects upon the synthesis of this enzyme.