Fig. 7.
Mutational analysis of the role of the secondary sugar-binding site that enhances trehalose binding to human mincle. (A–C) Purification of mutant proteins on invertase-Sepharose. Fractions eluted from the column were analyzed on parallel SDS–polyacrylamide gels that were stained with Coomassie blue or blotted and probed with alkaline phosphatase-conjugated avidin. (D–F) Comparison of binding of mutant proteins to trehalose and α-methyl glucoside using binding competition assay. Experimental data are represented by dots and curves obtained by nonlinear least-squares fitting are shown as solid lines. Mean ± SD for the ratios of the KI values derived from three replicate experiments are indicated.