Crystal Structure of Vaccinia Viral A27 Protein Reveals a Novel Structure Critical for Its Function and Complex Formation with A26 Protein
Figure 6
Immunoblot analysis of A27 proteins in transiently transfected-infected cells.
293T cells were infected with WRΔA27L virus and subsequently transfected with plasmids containing A27-wt, A27-TM-N, A27-TM-C, or A27-6A DNA. Lysates were harvested at 24 h post-infection and separated on 4% to 12% SDS-PAGE gels in reducing (+2ME) (A) or non-reducing (−2ME) conditions (B and C) for immunoblot analysis with anti-A27 (1∶1,000) (A & B) and anti-A26 (1∶5,000) (A & C) antibodies. The arrowheads labeled A27-mono, A27-di, and A27-tri represent A27 protein monomer, dimer, and trimer, respectively. The black and white arrows mark the 90-kDa and 70-kDa A26–A27 protein complexes, respectively. A26-containing bands appeared as doublets because of the incomplete formation of intramolecular disulfide bonding between C43 and C342 in the A26 protein [23].