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. 2011 Apr 1;286(21):18930–18937. doi: 10.1074/jbc.M111.234583

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Side of NH₂OH entry into light-activated Rho. Two types of Rho-containing proteoliposomes (prepared in either the absence or presence of NH₂OH) were exposed to light while they were maintained in NH₂OH-free buffers. Extracted retinoids were analyzed by HPLC. A, chromatograms show HPLC separation of at-RAL (peak 1) and its oxime (syn) (peak 2). Red traces correspond to proteoliposomes loaded with NH₂OH, and blue traces represent data obtained from the empty vesicles. Panel a, extraction with hexane; panel b, disruption of proteoliposomes with methanol prior to extraction; panel c, incubation with NH₂OH added after light exposure and 5 min prior to extraction. B, identification of detected retinoids. Peaks corresponding to at-RAL (left panel) and its oxime (syn) (right panel) were assigned based on the characteristic shapes and absorbance maxima of their UV-visible spectra. mAU, milliabsorbance units.