[Screening cellular proteins interacted with M2 protein of influenza A virus by coimmunoprecipitation].
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 specifically 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).


CONCLUSIONS
Interaction between Ataxin 10 and A/M2 would explain why influenza 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.
Li
Yaodong
Y
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. liyaodong@im.ac.cn
Guan
Zhenhong
Z
Yan
Jinghua
J
D000067736:Ataxin-10; D002460:Cell Line; D039642:Eukaryotic Initiation Factors / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D009980:Influenza A virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007251:Influenza, Human / Q000378:metabolism* / Q000821:virology; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D011485:Protein Binding; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000086082:Viroporin Proteins
D000067736:Ataxin-10; D039642:Eukaryotic Initiation Factors; D009419:Nerve Tissue Proteins; D014763:Viral Matrix Proteins; C503889:ATXN10 protein, human; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins