A protease of Bacteroides gingivalis degrades cell surface and matrix glycoproteins of cultured gingival fibroblasts and induces secretion of collagenase and plasminogen activator
A protease of Bacteroides gingivalis degrades cell surface and matrix glycoproteins of cultured gingival fibroblasts and induces secretion of collagenase and plasminogen activator
复制标题
牙龈拟杆菌的蛋白酶降解培养的牙龈成纤维细胞的细胞表面和基质糖蛋白，并诱导胶原酶和纤溶酶原激活剂的分泌
DOI：
10.1128/iai.57.1.213-218.1989
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发表时间：
1989
影响因子：
3.1
通讯作者：
Timo Sorsa
中科院分区：
文献类型：
--
作者：
V. Uitto;Hannu Larjava;Jyrki Heino;Timo Sorsa
为评价牙龈类杆菌对牙周细胞的直接作用，将牙龈类杆菌提取液或部分纯化的类胰酶作用于人牙龈成纤维细胞，用苯甲酰亚胺-琼脂糖层析法和S琼脂糖凝胶柱层析法进行分离纯化。用高碘酸盐-[~3H]硼氢化物标记技术结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)-荧光法对细胞表面糖蛋白的分析表明，纤维连接蛋白和其他一些高相对分子质量的细胞表面糖蛋白被35,000-MR(35K)牙龈假单胞菌蛋白酶降解。成纤维细胞培养物免疫染色显示细胞间基质纤维连接蛋白被35K酶降解。通过SDS-PAGE-免疫印迹技术检测反应产物，监测纤维连接蛋白的降解模式。与相应数量的胰腺胰蛋白酶相比，该酶能更快、更广泛地降解纤维连接蛋白。将细胞培养液与可溶性I型胶原在25℃下孵育，然后对反应产物进行SDS-PAGE-荧光分析，以检测成纤维细胞胶原酶的分泌。用酪蛋白-琼脂糖凝胶扩散平板法检测纤溶酶原激活物活性。用35K酶培养的成纤维细胞分泌较多的胶原酶和纤溶酶原激活剂。结果提示，牙周感染牙周白假单胞菌可引起宿主细胞表面结构的蛋白分解损伤。与此同时，牙龈假单胞菌可能会诱导细胞降解其细胞周围基质。
To assess the direct effects of Bacteroides gingivalis on periodontal cells, human gingival fibroblasts were cultured in the presence of B. gingivalis extracts or a trypsinlike enzyme partially purified from the bacteria by chromatography on benzamidine-Sepharose and Sephacryl S-200. Analysis of cell surface glycoproteins by the periodate-[3H]borohydride labeling technique combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-fluorography demonstrated that fibronectin and some other high-molecular-weight cell surface glycoproteins were degraded by a 35,000-Mr(35K) B. gingivalis protease. Immunostaining of the fibroblast cultures showed degradation of intercellular matrix fibronectin by the 35K protease. The pattern of fibronectin degradation was monitored by examining the reaction products with the SDS-PAGE-immunoblotting technique. The protease degraded fibronectin rapidly and more extensively than did corresponding amounts of pancreatic trypsin. Collagenase secretion by the fibroblasts was assayed by incubating cell culture medium with soluble type I [3H]collagen at 25 degrees C followed by SDS-PAGE-fluorography analysis of the reaction products. The medium was also assayed for plasminogen activator activity by using a casein-agarose diffusion plate assay. The fibroblasts cultured with the 35K protease secreted increased amounts of collagenase and plasminogen activator into the medium. The results suggest that periodontal infection by B. gingivalis causes proteolytic damage of the host cell surface structures. Concomitantly, B. gingivalis may induce the cells to degrade their pericellular matrix.
影响因子：
56.9
作者：
HOLT, SC;EBERSOLE, J;KORNMAN, KS
通讯作者：
KORNMAN, KS
DOI：
--
发表时间：
1983
期刊：
Collagen and related research
影响因子：
--
作者：
Narayanan,AS;Page,RC
通讯作者：
Page,RC