FIGURE 8.

Physical interaction of wild-type gp5 and gp5-C313S with trx. Separation of wt gp5/trx from free wt gp5 was performed on a hydroxyapatite chromatography in phosphate buffer as described under “Experimental Procedures.” Wild-type gp5 or gp5-C313S (5 μm) was premixed with trx (50 μm) and dialyzed against 500 ml of a buffer containing 10 mm potassium phosphate, pH 7.4, 1 mm DTT, 2 mm sodium citrate, and 10% glycerol. Dialyzed proteins were loaded onto the hydroxyapatite column using a FPLC system. Proteins were eluted with a 100-ml gradient from 10 to 250 mm potassium phosphate, pH 7.4. A, a sample containing wild-type gp5 and trx is shown. B, a sample containing gp5-C313S and trx is shown.