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Effect of replacements of amino acid residues outside the active site on difference spectra and apparent Kd values of the CYP46A1 mutants. Insets show the fitting of the binding curves. BIC binding properties of the CYP46A1 wild type (WT) are shown for comparison. Titrations were carried with racBIC, and S and R isomers as described under “Experimental Procedures.” The results represent the average of triplicate measurements ± S.D.
