Figure 6.
Characterization of WT, Y75C, and I99C β-tryptases in complex with murine B12 Fab. A, size-exclusion chromatography of B12 Fab in complex with WT, Y75C, and I99C β-tryptases was carried out as in Fig. 2 on an S200GL column in SEC buffer. Chromatograms are colored as follows: WT tryptase tetramer is colored black (Peak 3; Ve = 13.0 ml); complexes of B12 Fab with WT are green (Peak 2; Ve = 14.3 ml), Y75C is blue (Peak 4; Ve = 12.1 ml), and I99C is red (Peak 5; Ve = 12.1 ml). Excess free B12 Fab in Peaks 1a,b,c all have a similar Ve = 15.7 ml. B, fractions of main peaks were analyzed by SDS-PAGE under nonreducing conditions.