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Figure 3. Design strategy of rhodopsin-based ICCRs and functional coupling.(A) Sequence alignment of the fusion area between GPCRs and Kir6.2ΔN25. Alignment of GPCR C-terminal sequences were based on the presence of the H8 Helix. (B) Percent change in current for each construct in response to 10 µM all-trans-retinal or light (after 11-cis retinal incubation). Oocytes were co-injected with the specified Ops-Kir6.2 and TMD0, a SUR transmembrane domain. Numbers below bars indicate the number of oocytes tested. The changes induced by all-trans-retinal and light were statistically significant for Ops-K−16–25 (Student t-test; P<0.04 & P<0.0001, respectively), but not for Ops-K0–25 (P>0.4 & P>0.05, respectively). (C) Representative TEVC recordings for each construct in the case of photoactivation. Yellow bar represents oocyte illumination. Blue bar corresponds to Ba2+ application at 3 mM. Dashed line indicates the Ba2+-sensitive current baseline. (D) Opsin ability to activate G proteins within the fusion Ops-Kir6.2. Both constructs were co-expressed with Kir3.1* and change in current was measured in response to 10 µM all-trans-retinal. All measurements were done at −50 mV. Numbers above bars indicate the number of oocytes tested. Image published in: Caro LN et al. (2012) Image reproduced on Xenbase with permission of the publisher and the copyright holder. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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