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Penzkofer, Alfons ; Silapetere, Arita ; Hegemann, Peter

Photocycle Dynamics of the Archaerhodopsin 3 Based Fluorescent Voltage Sensor QuasAr1

Penzkofer, Alfons, Silapetere, Arita and Hegemann, Peter (2019) Photocycle Dynamics of the Archaerhodopsin 3 Based Fluorescent Voltage Sensor QuasAr1. International Journal of Molecular Sciences 21 (160), pp. 1-22.

Date of publication of this fulltext: 09 Jan 2020 13:33
Article
DOI to cite this document: 10.5283/epub.41351


Abstract

The retinal photocycle dynamics of the fluorescent voltage sensor QuasAr1 (Archaerhodopsin 3 P60S-T80S-D95H-D106H-F161V mutant from Halorubrum sodomense) in pH 8 Tris buffer was studied. The samples were photoexcited to the first absorption band of the protonated retinal Schiff base (PRSB) Ret_580 (absorption maximum at lambda(max) approximate to 580 nm), and the retinal Schiff base ...

The retinal photocycle dynamics of the fluorescent voltage sensor QuasAr1 (Archaerhodopsin 3 P60S-T80S-D95H-D106H-F161V mutant from Halorubrum sodomense) in pH 8 Tris buffer was studied. The samples were photoexcited to the first absorption band of the protonated retinal Schiff base (PRSB) Ret_580 (absorption maximum at lambda(max) approximate to 580 nm), and the retinal Schiff base photoisomerization and protonation state changes were followed by absorption spectra recordings during light exposure and after light exposure. Ret_580 turned out to be composed of two protonated retinal Schiff base isomers, namely Ret_580(I) and Ret_580(II). Photoexcitation of Ret_580(I) resulted in barrier-involved isomerization to Ret_540 (quantum yield approximate to 0.056) and subsequent retinal proton release leading to Ret_410 deprotonated retinal Schiff base (RSB). In the dark, Ret_410 partially recovered to Ret_580(I) and partially stabilized to irreversible Ret_400 due to apoprotein restructuring (Ret_410 lifetime approximate to 2 h). Photoexcitation of Ret_580(II) resulted in barrier-involved isomerization to Ret_640 (quantum yield approximate to 0.00135) and subsequent deprotonation to Ret_370 (RSB). In the dark, Ret_370 partially recovered to Ret_580(II) and partially stabilized to irreversible Ret_350 due to apoprotein restructuring (Ret_370 lifetime approximate to 10 h). Photocycle schemes and reaction coordinate diagrams for Ret_580(I) and Ret_580(II) were developed and photocyle parameters were determined.



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Details

Item typeArticle
Journal or Publication TitleInternational Journal of Molecular Sciences
Publisher:MDPI
Place of Publication:BASEL
Volume:21
Number of Issue or Book Chapter:160
Page Range:pp. 1-22
Date25 December 2019
InstitutionsPhysics > Institute of Experimental and Applied Physics
Physics > Institute of Experimental and Applied Physics > Alumni or Retired Professors > Group Alfons Penzkofer
Physics > Institute of Experimental and Applied Physics > Alumni or Retired Professors
Identification Number
ValueType
10.3390/ijms21010160DOI
KeywordsALL-OPTICAL ELECTROPHYSIOLOGY; ACTION-POTENTIALS; SCHIFF-BASE; ABSORPTION; PROTEIN; NEURONS; DEPROTONATION; RHODOPSIN; QuasAr1; Archaerhodopsin 3; genetically encoded fluorescent voltage sensor; absorption spectroscopic characterization; fluorescence studies; photocycle dynamics; photoisomerization; deprotonation; reprotonation
Dewey Decimal Classification500 Science > 530 Physics
StatusPublished
RefereedYes, this version has been refereed
Created at the University of RegensburgYes
URN of the UB Regensburgurn:nbn:de:bvb:355-epub-413517
Item ID41351

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