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. 2022 Aug 4;12(8):1076. doi: 10.3390/biom12081076

Figure 11.

Figure 11

Reconstitution and functional characterization of CYP450 2B4 in differently charged polymer-nanodiscs and DPC micelles: (a) Schematic showing CYP450 with heme coordination spheres of the CO-bound state reconstituted in an SMA-QA:DMPC nanodisc. (b) UV-vis absorption spectra of CYP450 reconstituted in different SMA polymer-nanodiscs or in DPC micelles in its ferric state (left column) and in a ferrous state in complex with CO (right column). UV-vis absorption spectra of a positively-charged CYP450 reconstituted in negatively-charged SMA-EA nanodiscs: (c) in the presence of the indicated NaCl concentrations and (d) the ferrous-CO complex (d). UV-vis spectra of a positively-charged CYP450 reconstituted in positively-charged SMA-QA-based DMPC nanodiscs and (e) in the presence of NaCl and (f) the ferrous-CO complex. These results demonstrate the importance of a membrane mimetic and polymer charge in nanodisc for the functional reconstitution of membrane proteins. The inactive CYP450 (i.e., P420) in the presence of DPC detergent or an anionic-polymer (like SMA or SMA-EA) is undesirable. The positively-charged SMA-QA retains the functional form of CYP450. This Figure and caption are adopted from reference [199].