EXPRESSION OF A METALLOPROTEINASE THAT DEGRADES NATIVE TYPE-V COLLAGEN AND DENATURED COLLAGENS BY CULTURED HUMAN ALVEOLAR MACROPHAGES
EXPRESSION OF A METALLOPROTEINASE THAT DEGRADES NATIVE TYPE-V COLLAGEN AND DENATURED COLLAGENS BY CULTURED HUMAN ALVEOLAR MACROPHAGES
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DOI：
10.1172/jci113253
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发表时间：
1987-12-01
影响因子：
15.9
通讯作者：
KANG, AH
中科院分区：
文献类型：
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作者：
HIBBS, MS;HOIDAL, JR;KANG, AH
从正常对照组和吸烟者的支气管肺泡灌洗获得的人肺泡巨噬细胞在体外分泌一种中性蛋白酶，降解变性胶原。在培养3-5 d后检测到蛋白酶的最佳表达。在用0.5 μ g/ml放线菌酮处理的培养物的培养基中不能检测到蛋白酶。明胶酶的Mr为90，000，与人中性粒细胞明胶酶具有免疫交叉反应性。当新合成的35 S-蛋氨酸标记的蛋白质进行了分析，蛋白酶似乎是肺泡巨噬细胞的主要分泌产物。在明胶-琼脂糖凝胶上层析得到主要由90，000-mol-wt蛋白酶组成的单一活性峰。明胶琼脂糖凝胶纯化的材料中的蛋白水解活性被EDTA和1，10-菲咯啉抑制，但不被N-乙基马来酰亚胺或苯基甲磺酰氟，表明该蛋白酶是金属蛋白酶。部分纯化的材料还能够降解天然V型胶原蛋白，并且在中性粒细胞明胶酶抗体的存在下，这种降解被抑制。这些数据表明，人类肺泡巨噬细胞在文化阐述了一种金属蛋白酶，降解天然V型胶原和变性胶原。
Human pulmonary alveolar macrophages obtained by bronchoalveolar lavage from both normal controls and smokers secreted in vitro a neutral proteinase that degraded denatured collagens. Optimal expression of the proteinase was detected after 3-5 d of culture. The proteinase could not be detected in the media of cultures that had been treated with 0.5 .mu.g/ml of cycloheximide. The gelatinase had an Mr of 90,000 and was immunologically cross-reactive with human neutrophil gelatinase. When newly synthesized 35S-methionine-labeled proteins were analyzed, the proteinase appeared to be a major secretion product of alveolar macrophages. Chromatography on gelatin-Sepharose gave a single peak of activity that was predominantly composed of the 90,000-mol-wt proteinase. The proteolytic activity in the gelatin-Sepharose-purified material was inhibited by EDTA and 1,10-phenanthroline, but not by N-ethylmaleimide or phenylmethanesulfonyl fluoride, indicating that the proteinase was a metalloproteinase. The partially purified material was also capable of degrading native type V collagen and this degradation was inhibited in the presence of an antibody to neutrophil gelatinase. The data suggest that human alveolar macrophages in culture elaborate a metalloproteinase that degrades both native type V collagen and denatured collagens.