Comparison of the molecular properties of retinitis pigmentosa P23H and N15S amino acid replacements in rhodopsin - Fig 2

A. Overlay of absorbance spectra of rhodopsin proteins expressed in COS-1 cells and purified using 1D4-affinity chromatography. Proteins were eluted in phosphate buffered saline in order to elute both folded and misfolded portions. B. P23H rhodopsin obtained from stable cell line P23H and eluted in 2 mM sodium phosphate pH 6, which only elutes the folded fraction. C. Rhodopsin stability measured by loss of 500 nm chromophore over time at 37°C. Absorbance spectra of WT, N15S and P23H rhodopsin expressed in the presence of 9-cis retinal at 37°C were measured over time, and the decrease in absorbance at 500 nm was expressed at percent of intact structure. D. Rhodopsin Meta-II decay monitored by the use of fluorescence spectroscopy. Fluorescence spectra of rhodopsin Meta-II decay of WT, N15S and P23H rhodopsin expressed in the presence of 9-cis retinal.