Glasgow J E, Bagdasarian A, Colman R W
J Lab Clin Med. 1982 Jan;99(1):108-17.
Alpha 1 protease inhibitor antigen was identified in the culture medium of the human ascites hepatoma cell line SK-HEP-1. Trypsin inhibitory activity and alpha 1 Pl antigen accumulated in serum-free medium concomitantly over a period of several days. Radioactive alpha 1 Pl antigen was detected in conditioned medium from cultures supplemented with 35S-L-methionine, indicating a synthesis and release of the protein. Alpha 1 Pl antigen in conditioned medium appeared to be antigenically identical to that in human plasma, and the newly synthesized (radiolabeled) antigen co-migrated with plasma, alpha 1 Pl after immunoelectrophoresis or SDS-polyacrylamide gel electrophoresis. Moreover, evidence is presented that the synthesized inhibitor exhibits functional activity, since the 35S-labeled alpha 1 Pl in conditioned medium complexes with trypsin. We conclude that SK-HEP-1 cells in culture produce functionally active alpha 1 Pl which may be identical to that in plasma.
在人腹水肝癌细胞系SK-HEP-1的培养基中鉴定出α1蛋白酶抑制剂抗原。在无血清培养基中,胰蛋白酶抑制活性和α1抗胰蛋白酶(α1Pl)抗原在数天内同时积累。在添加了35S-L-甲硫氨酸的培养物的条件培养基中检测到放射性α1Pl抗原,表明该蛋白的合成与释放。条件培养基中的α1Pl抗原在抗原性上似乎与人血浆中的相同,并且新合成的(放射性标记的)抗原在免疫电泳或SDS-聚丙烯酰胺凝胶电泳后与血浆α1Pl共迁移。此外,有证据表明合成的抑制剂具有功能活性,因为条件培养基中35S标记的α1Pl与胰蛋白酶形成复合物。我们得出结论,培养的SK-HEP-1细胞产生功能活性的α1Pl,其可能与血浆中的α1Pl相同。