Purification and Characterization of a Human Mx Protein

Abstract

Human interferon-beta (IFN-beta) induces in human embryonic foreskin fibroblasts a cytoplasmic protein with antigenic similarities to mouse Mx protein, a nuclear protein implicated in inhibition of influenza virus replication. The human protein was purified to virtual homogeneity by immunoaffinity chromatography using a monoclonal antibody to mouse Mx protein. The purified protein has an apparent Mr of 78,000 and displays a strong tendency to self-aggregate. It can be resolved on two-dimensional gels into four spots with pIs between 6.0 and 6.2, each of which reacts with antibodies to mouse Mx protein. The partial amino-terminal sequence was determined for the affinity-purified protein. Cytoplasmic microinjection of the affinity-purified protein does not lead to efficient protection against infection with influenza virus. Cytoplasmic microinjection of the monoclonal Mx antibody, which increases suceptibility of IFN-treated mouse Mx cells to influenza virus, does not alter the viral susceptibility of IFN-treated human cells. These results suggest that, unlike the mouse Mx protein, the human Mx protein studied in this communication may not be sufficient to confer to cells a high degree of protection against influenza virus.