Penzkofer, Alfons and Silapetere, Arita and Hegemann, Peter (2021) Photocycle dynamics of the Archaerhodopsin 3 based fluorescent voltage sensor Archon2. JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY, 225: 112331. ISSN 1011-1344, 1873-2682
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The retinal photocycle dynamics of the fluorescent voltage sensor Archon2 in pH 8 Tris buffer was studied. Archon2 is a mutant of Archaerhodopsin 3 (Arch) from Halorubrum sodomense obtained by a robotic multidimensional directed evolution approach (Archon2 = Arch T56P-P60S-T80P-D95H-T99S-T116I-F161V-T183IL197I-A225C). The samples were photo-excited to the first absorption band of the protonated retinal Schiff base (PRSB) Ret_586 (absorption maximum at lambda(max) = 586 nm, excitation wavelengths lambda(exc) = 590 nm and 632.8 nm). The photocycle dynamics were studied by recording absorption spectra during light exposure and after light exposure. Ret_586 photoisomerized to Ret_535 (main component) and Ret_485 (minor component). Ret_535 backward photoisomerized to Ret_586 in light-adapted state (named Ret_586(la)) and partly deprotonated to neutral retinal Schiff base (RSB) Ret_372 in light adapted state (named Ret_372(la), same isomer form as Ret_535). After excitation light switch-off Ret_372(la) recovered to Ret_372 in dark-adapted state (Ret_372(da)) which slowly re-protonated to Ret_535, and Ret_535 slowly isomerized back to Ret_586 in dark-adapted state (Ret_586(da)). Photocycle schemes and reaction coordinate diagrams are developed and photocycle parameters are determined.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ALL-OPTICAL ELECTROPHYSIOLOGY; SENSITIVE DYES; SCHIFF-BASE; RHODOPSIN; ABSORPTION; DEPROTONATION; INTERSECTION; MECHANISM; LIFETIME; PROTEIN; Archon2; Archaerhodopsin 3; Genetically encoded voltage sensor (GEVI); Absorption spectroscopic studies; Photocycle dynamics; Photoisomerization; Thermal activated isomerization; Thermal activated deprotonation; Thermal activated re-protonation |
| Subjects: | 500 Science > 530 Physics |
| Divisions: | Physics > Institute of Experimental and Applied Physics > Alumni or Retired Professors > Group Alfons Penzkofer |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 10 Aug 2022 10:51 |
| Last Modified: | 10 Aug 2022 10:51 |
| URI: | https://pred.uni-regensburg.de/id/eprint/46419 |
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