Baculoviral expression of a natural inhibitor of the human insulin receptor tyrosine kinase.

We had earlier reported that the human serum alpha 2-HS glycoprotein (1) is a physiological and specific inhibitor of the human insulin receptor tyrosine kinase (IR-TK). We have now expressed this human protein in the baculoviral expression system using the Sf-9 and High Five insect cells. The protein was optimally expressed at 72 h post infection. alpha 2-HSGbac completely inhibited the insulin-stimulated autophosphorylation and TK activity of partially purified IR preparations. It also abolished insulin-induced DNA synthesis in the H-35 rat hepatoma cell line. The effective concentration of the baculoviral derived alpha 2-HSG necessary for inhibiting IR-TK activity was significantly lower than that of the protein purified from human plasma.