Expression of Soluble Recombinant Integrin Alpha V and Beta 3 from SF21 Cells with Baculovirus Expression Vector for Targeted Screening of Integrin Antagonists

摘要

We examined production of membrane domain-truncated integrin alpha_v and beta_3 in an insect cell line, SF21, by using baculovirus expression vector for screening of new ligands interacted with integrin alpha_v beta_3. Molecular cloning of the integrin receptors was carried out by truncation of transmembrane and cytoplasmic domains. Extracellular domains of integrin alpha_v and beta_3 were expressed in SF 21 cells and purified by His-tag affinity column chromatography with high purity. Activities ofpurified recombinant integrin alpha_v and beta_3 were confirmed by solid phase integrin binding assay via direct interaction method of integrin alpha_v beta_3-vitronectin on protein microarray chip. Taken together, these data suggests that transmembrane-deleted integrin alpha_v and beta_3 can be massively produced in Sf21 insect cell expression system with full biological activity and applied to screen novel antagonists from different libraries such as peptide, chemical compound, and natural product libraries.