Identification of substrate molecules on the host immune cells of bacterial proteases by using radioisotopes and mass spectrometry.

来自健康志愿者外周血单核细胞的中性粒细胞进行细胞表面放射性碘标记和细菌蛋白酶处理。中性粒细胞非常脆弱，很容易崩溃，很难将细胞表面的蛋白质与细胞质中的蛋白质分开。因此，正在寻找适合中性粒细胞的放射性标记和蛋白水解的条件。为了从凝胶中回收放射性蛋白质，通过使用干净的冲头从夹在玻璃纸片中的干燥凝胶中冲压出蛋白质斑点是令人满意的。为了保护那些不使用放射性同位素的人，有必要防止真空泵排出的带有微量放射性的废气在实验室中扩散。在这项研究中，确定了金黄色葡萄球菌葡萄球菌蛋白酶对纤维蛋白原和胶原蛋白的新影响，这是金黄色葡萄球菌对凝血受损和组织破坏的毒力活动的原因。

Neutrophils from peripheral blood mononuclear cells of healthy volunteers were subjected to cell-surface radioactive-iodine labeling and to bacterial protease treatment. Neutrophils were so fragile and easily collapsed that it was difficult to separate proteins on the cell surface from those in the cytosol. Therefore, searching the conditions suitable for the radiolabeling and proteolysis for the neutrophils are underway. To recover the radioactive proteins from the gels, it was satisfactory to punch out the protein spots from dried gels sandwiched by cellophane sheets by using clean punches. It was necessary to prevent the exhaust air with trace amount of radioactivity from the vacuum pumps from diffusing in the laboratory to take care those people who do not use radioisotopes.In this study, new effects of Staphylococcus aureus staphopains on fibrinogen and collagen were identified that account for the virulence activities of S aureus for impaired clotting and tissue destruction.