Screening cellular proteins interacted with M2 protein of influenza A virus by Coimmunoprecipitation

Abstract:

Abstract: [Objective] To screen cellular protein interacted with influenza A M2 protein(A/M2). [Methods] we cloned A/M2 gene fragment into pCAGGS-CFlag vector, and the resulting plasmid was transfected into human embryonic kidney (HEK) 293T cells.The recombinant Flag fusion protein,A/M2-Flag was absorbed specificly by Anti-Flag Monoclonal Antibody M2-Conjugated Agarose beads, we loaded the beads on 12% SDS-PAGE after we washed it with lysis buffer. Silver staining of the gel revealed that several proteins were co-purified with A/M2.To identify the proteins,we excised the protein bands and analysed them by mass spectroscopic sequencing. [Results] We got two kinds of proteins, ataxin 10 and eukaryotic initiation factors(eIFs). [Conclusion] Interaction between Ataxin 10 and A/M2 would explain why inflenza virus infection or influenza vaccine innoculation causes acute cerebellar ataxia. A/M2 interacting with eIFs would imply that A/M2 is involved in the regulation of influenza virus protein synthesis.