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. 1999 Mar 2;96(5):1932–1935. doi: 10.1073/pnas.96.5.1932

Figure 2.

Figure 2

Characterization of the expressed mutants C110A, C110Y, and C110F. (A) UV–vis absorption spectra of the mutants isolated after treatment of COS-1 cells with 11-cis-retinal. As indicated, COS-1 cells expressing the opsin from C110A were treated with 5 and 50 μM retinal; the opsins from C110F and C110Y were treated with 50 μM retinal (Methods). Elution of the proteins from 1D4-Sepharose was with Buffer A containing 0.05% DM and 100 μM peptide. (B) I, C110A: Elution profile from 1D4-Sepharose of the in vivo retinal-treated (50 μM) C110A mutant opsin. Elution of the first eight fractions (300 μl each) was with Buffer J; the following seven fractions were eluted with Buffer K, which contains salt. II: Bleaching behavior of the fraction (no. 2) eluted with Buffer J (no salt) in B. The UV–vis spectrum showed an A280/A500 ratio of 3.9, indicating there was contamination from unconstituted opsin.