Figure 4
The characteristics of the R264H mutant in solution. (a) A thermal shift assay shows the Tm of the R264H mutant (dimer and tetramer) compared with the wild type. (b) Gel-filtration profiles of wild-type MATα1 (blue) and the R264H mutant (green) are shown by the absorbance at A280. The vertical markers (1–5) represent the elution volumes of standard proteins (GE Healthcare): ferritin (1; 440 kDa), aldolase (2; 158 kDa), conalbumin (3; 75 kDa), ovalbumin (4; 43 kDa) and ribonuclease (5; 13.7 kDa). (c) Native PAGE and SDS–PAGE gels of the MATα1 R264H–MATβV1 fraction are presented with those of R264H MATα1 and MATβV1 on their own. (d) Gel-filtration profiles of the R264H mutant with or without SCR0911 incubation are shown. (e) SAMe production by the R264H mutant at pH 6.0 and 7.5 compared with the wild type. Data are the mean ± SEM (n = 3). |