Miszczuk-Jamska B, Merten M, Guy-Crotte O, Amouric M, Clemente F, Schoumacher R A, Figarella C
Groupe de Recherche sur les Glandes Exocrines, Marseille, France.
FEBS Lett. 1991 Dec 9;294(3):175-8. doi: 10.1016/0014-5793(91)80662-m.
Proteins with trypsin-like immunoreactivity (first detected by a specific immunoenzymatic assay) were isolated from CAPAN-1 and CFPAC-1 cell culture-conditioned media by chromatography on an immunoadsorbent prepared with a polyclonal antibody directed against trypsin 1. The adsorbed proteins were devoid of free trypsin activity but trypsin activity was present after enterokinase activation demonstrating that the immunoreactive trypsin present in cell supernatants corresponds to trypsinogens. When characterised by Western blotting using a monoclonal antibody directed against human trypsin 1 two protein bands corresponding to trypsinogen 1 (23 kDa) and trypsinogen 2 (25 kDa) gave a positive reaction. These results demonstrate the presence of trypsinogens 1 and 2 in CAPAN-1 and CFPAC-1 cells and in their culture-conditioned media.
具有类胰蛋白酶免疫反应性的蛋白质(首先通过特定免疫酶测定法检测到),通过在由针对胰蛋白酶1的多克隆抗体制备的免疫吸附剂上进行色谱分离,从CAPAN-1和CFPAC-1细胞培养条件培养基中分离出来。吸附的蛋白质没有游离的胰蛋白酶活性,但在肠激酶激活后存在胰蛋白酶活性,这表明细胞上清液中存在的免疫反应性胰蛋白酶对应于胰蛋白酶原。当使用针对人胰蛋白酶1的单克隆抗体通过蛋白质印迹法进行表征时,两条对应于胰蛋白酶原1(23 kDa)和胰蛋白酶原2(25 kDa)的蛋白带呈阳性反应。这些结果证明了CAPAN-1和CFPAC-1细胞及其培养条件培养基中存在胰蛋白酶原1和2。