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. Author manuscript; available in PMC: 2008 Oct 6.
Published in final edited form as: Biochemistry. 2006 Jan 24;45(3):746–754. doi: 10.1021/bi050870z

Figure 5.

Figure 5

Photolabeling of cytochrome c oxidase by bis-(AzC12)-CL or SAND-gly-CL. Panel A shows photolabeling of CL-free (filled bars) and CL-containing CcO (open bars) by bis-(AzC12)-CL. Panel B shows photolabeling of CL-free (filled bars) and CL-containing CcO (open bars) by SAND-gly-CL. Percent photolabeling was calculated from the decreased area of the RP-HPLC elution peak for enzyme that had been reconstituted with either bis-(AzC12)-CL or SAND-gly-CL, purified by HiTrap Q ion exchange chromatography to remove free ligand, and exposed or not exposed to intense visible light. Average values and standard deviations were calculated from three separate paired experiments. The percent labeling of subunit VIa is based upon the total decreased HPLC yield of subunit VIa plus Ac-VIa. In each analysis, 100 μg (0.5 nmol) of CcO was loaded on the RP-column, and the subunit content was quantitatively determined according to Liu et al. (30).