Figure 5.
Figure 5. Analysis of acidic pH–induced trimer formation with WT and mutant RSPs by rate zonal gradient centrifugation. (A–D) RSPs were incubated for 10 min at pH 5.4 (solid line) or 8.0 (dotted line), back-neutralized, solubilized with 1% Triton X-100, and analyzed by sedimentation in 7–20% sucrose gradients containing 0.1% Triton X-100. The gradients were fractionated, and the amount of E protein in each fraction was determined by a quantitative four-layer ELISA. The sedimentation direction is from left to right. Peak of the E dimer, fraction 8; peak of the E trimer, fraction 10.

Analysis of acidic pH–induced trimer formation with WT and mutant RSPs by rate zonal gradient centrifugation. (A–D) RSPs were incubated for 10 min at pH 5.4 (solid line) or 8.0 (dotted line), back-neutralized, solubilized with 1% Triton X-100, and analyzed by sedimentation in 7–20% sucrose gradients containing 0.1% Triton X-100. The gradients were fractionated, and the amount of E protein in each fraction was determined by a quantitative four-layer ELISA. The sedimentation direction is from left to right. Peak of the E dimer, fraction 8; peak of the E trimer, fraction 10.

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