DISPASE, A NEUTRAL PROTEASE FROM BACILLUS-POLYMYXA, IS A POWERFUL FIBRONECTINASE AND TYPE-IV COLLAGENASE
DISPASE, A NEUTRAL PROTEASE FROM BACILLUS-POLYMYXA, IS A POWERFUL FIBRONECTINASE AND TYPE-IV COLLAGENASE
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DOI：
10.1111/1523-1747.ep12277593
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发表时间：
1989-08-01
影响因子：
6.5
通讯作者：
KUKLINSKA, E
中科院分区：
文献类型：
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作者：
STENN, KS;LINK, R;KUKLINSKA, E
分散酶是从多粘芽孢杆菌的培养物中分离的中性蛋白酶，已被证明是一种快速、有效但温和的试剂，用于分离培养物中的完整表皮与真皮以及完整上皮片与基质。在这两种情况下，它通过切割基底膜区而实现分离，同时保留上皮细胞的活力。因为不知道分散酶在基底膜区切割什么或在哪里，我们建立了研究来定义其底物特异性。使用纯化的基底膜成分和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，我们表明，Dispase裂解纤连蛋白和IV型胶原，但不层粘连蛋白，V型胶原，血清白蛋白，或转铁蛋白。分散酶对胶原蛋白的作用似乎对IV型胶原蛋白具有选择性，因为会形成几种稳定的降解产物，而酶仅最低限度地降解I型胶原蛋白。在新生儿皮肤中，如电子显微镜所见，Dispase去除了富含IV型胶原的致密层，但保留了锚定原纤维（已知含有VII型胶原的结构）和表皮细胞。因为它的作用是如此的选择性，它表明Dispase可以作为一个强大的工具，解剖上皮间充质相互作用。
Dispase, a neutral protease isolated from culture filtrates of Bacillus polymyxa, has proven to be a rapid, effective, but gentle agent for separating intact epidermis from the dermis and intact epithelial sheets in culture from the substratum. In both cases it effects separation by cleaving the basement membrane zone region while preserving the viability of the epithelial cells. Because it is not known what or where in the basement membrane zone Dispase cleaves, we set up studies to define its substrate specificity. Using purified basement membrane components and sodium dodecyl sulfate-polyacrylamide gel electrophoresis we show that Dispase cleaves fibronectin and type IV collagen, but not laminin, type V collagen, serum albumin, or transferrin. The action of Dispase on collagen appears to be selective for type IV collagen in that several stable degradation products are formed, whereas the enzyme degrades type I collagen only minimally. In newborn human skin, as seen by electron microscopy, Dispase removes the lamina densa, rich in type IV collagen, but preserves the anchoring fibrils (structures known to contain type VII collagen) and the epidermal cells. Because its action is so selective, it suggests that Dispase can serve as a powerful tool for dissecting epithelial-mesenchymal interactions.