Grenier D, Uitto V J, McBride B C
Departement de Santé Buccale, Université de Montréal, Québec, Canada.
Infect Immun. 1990 Feb;58(2):347-51. doi: 10.1128/iai.58.2.347-351.1990.
A number of immunological methods were used to localize a cell-associated Treponema denticola chymotrypsinlike protease. Indirect immunofluorescence staining, immunogold labeling, and an enzyme-linked immunosorbent assay all indicated that the protease was attached to the outside of the cell envelope. The invasive capability of T. denticola was evaluated by following the degradation of a reconstituted basement membrane material (Matrigel) and the release of spirochetes from the gel. Under conditions where the chymotrypsinlike activity was increased, more spirochetes migrated from the gel. Protease inhibitors strongly reduced the number of cells that moved out of the gel. The purified chymotrypsinlike protease degraded the basement membrane components type IV collagen, laminin, and fibronectin. The study suggests that the T. denticola chymotrypsinlike protease may play an important role in the invasion and destruction of basement membrane.
运用了多种免疫学方法来定位一种与细胞相关的齿垢密螺旋体类胰凝乳蛋白酶。间接免疫荧光染色、免疫金标记和酶联免疫吸附测定均表明该蛋白酶附着于细胞膜外表面。通过追踪重组基底膜材料(基质胶)的降解以及螺旋体从凝胶中的释放情况,评估了齿垢密螺旋体的侵袭能力。在类胰凝乳蛋白酶活性增强的条件下,更多螺旋体从凝胶中迁移出来。蛋白酶抑制剂显著减少了从凝胶中移出的细胞数量。纯化的类胰凝乳蛋白酶降解了基底膜成分IV型胶原蛋白、层粘连蛋白和纤连蛋白。该研究表明,齿垢密螺旋体类胰凝乳蛋白酶可能在基底膜的侵袭和破坏中起重要作用。