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. 2005 Sep 6;102(38):13658–13663. doi: 10.1073/pnas.0504167102

Fig. 4.

Fig. 4.

Effect of mutation of iron-coordinating residues on RPE65 activity and expression. (A) Mutation of iron-coordinating residues abolishes isomerase activity. Synthesis of 11-cis-retinol is normalized to RPE65 immunoreactivity quantified by densitometry. Relative activities of mutants are compared with wild type, expressed as the mean and SD of three determinations. (B) Effect of histidine and glutamate mutations on RPE65 expression. Equivalent volumes of whole-cell lysates of transfections with pVitro2/RPE65+CRALBP and pVitro3/LRAT+RDH5 constructs analyzed by immunoblot using RPE65 (Upper) and CRALBP (Lower) antibodies. The expression level of all mutants except H180A was lower than wild type. Lane 1, untransfected control; lane 2, wild-type RPE65; lane 3, H180A; lane 4, H241A; lane 5, H313A; lane 6, H527A; lane 7, E417A. The lower band in Upper is a nonspecific reactant present in both untransfected and transfected 293-F cells.