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. 2009 Mar 20;284(12):8064–8072. doi: 10.1074/jbc.M808942200

FIGURE 2.

FIGURE 2.

Effect of conservative mutations of residues Asp-431 and Asp-391 on pVP2/VP2 proteolytic maturation. A, Western blot analysis of proteins expressed in cells infected with rVV VT7LacOI/POLY, VT7LacOI/D431N, or VT7LacOI/D391N, respectively. Infected cultures were harvested at 24, 48, and 72 h pi, respectively. The corresponding extracts were subjected to SDS-PAGE and Western blot using anti-VP2 serum. B, QM7 cells infected with rVV VT7LacOI/POLY, VT7LacOI/D431N, or VT7LacOI/D391N were metabolically pulse-labeled with [35S]Met for 1 h at 19 hpi (hpi). The radioactivity was chased with by adding fresh medium containing an excess of cold methionine. Cultures were harvested at 0, 24, 48, and 72 h post-labeling, and the corresponding extracts were immunoprecipitated using anti-VP2 serum. The resulting samples were subjected to SDS-PAGE followed by autoradiography. Molecular weight markers (expressed in kDa, left) and bands corresponding to proteins pVP2 and VP2 (right) are indicated.