Proteorhodopsins are heptahelical membrane proteins which function as light-driven proton pumps. They use all-trans-retinal A1 as a ligand and chromophore and absorb visible light (520–540 nm). In the present paper, we describe modulation of the absorbance band of the proteorhodopsin from Monterey Bay SAR 86 gammaproteobacteria (PR), its red-shifted double mutant PR-D212N/F234S (PR-DNFS) and Gloeobacter rhodopsin (GR). This was approached using three analogues of all-trans-retinal A1, which differ in their electronic and conformational properties: all-trans-6,7-s-trans-locked retinal A1, all-trans-phenyl-retinal A1 and all-trans-retinal A2. We further probed the effect of these retinal analogues on the proton pump activity of the proteorhodopsins. Our results indicate that, whereas the constraints of the retinal-binding pocket differ for the proteorhodopsins, at least two of the retinal analogues are capable of shifting the absorbance bands of the pigments either bathochromically or hypsochromically, while maintaining their proton pump activity. Furthermore, the shifts implemented by the analogues add up to the shift induced by the double mutation in PR-DNFS. This type of chromophore substitution may present attractive applications in the field of optogenetics, towards increasing the flexibility of optogenetic tools or for membrane potential probes.
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Research Article|
April 02 2015
Modulation of spectral properties and pump activity of proteorhodopsins by retinal analogues
Srividya Ganapathy;
Srividya Ganapathy
1
*Department of Biophysical Organic Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands
†BioSolar Cells, P.O. Box 98, 6700 AB Wageningen, The Netherlands
1Correspondence may be addressed to either of these authors (email s.ganapathy@chem.leidenuniv.nl or w.j.de.grip@umail.leidenuniv.nl).
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Odette Bécheau;
Odette Bécheau
*Department of Biophysical Organic Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands
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Hanka Venselaar;
Hanka Venselaar
‡Centre for Molecular and Biomolecular Informatics, Radboud University Medical Center, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands
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Siebren Frölich;
Siebren Frölich
*Department of Biophysical Organic Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands
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Jeroen B. van der Steen;
Jeroen B. van der Steen
§Swammerdam Institute for Life Sciences, University of Amsterdam, P.O. Box 94215, 1090 GE Amsterdam, The Netherlands
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Que Chen;
Que Chen
§Swammerdam Institute for Life Sciences, University of Amsterdam, P.O. Box 94215, 1090 GE Amsterdam, The Netherlands
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Sarah Radwan;
Sarah Radwan
*Department of Biophysical Organic Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands
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Johan Lugtenburg;
Johan Lugtenburg
*Department of Biophysical Organic Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands
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Klaas J. Hellingwerf;
Klaas J. Hellingwerf
§Swammerdam Institute for Life Sciences, University of Amsterdam, P.O. Box 94215, 1090 GE Amsterdam, The Netherlands
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Huub J. M. de Groot;
Huub J. M. de Groot
*Department of Biophysical Organic Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands
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Willem J. de Grip
Willem J. de Grip
1
*Department of Biophysical Organic Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands
1Correspondence may be addressed to either of these authors (email s.ganapathy@chem.leidenuniv.nl or w.j.de.grip@umail.leidenuniv.nl).
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Publisher: Portland Press Ltd
Received:
September 30 2014
Revision Received:
January 27 2015
Accepted:
February 06 2015
Accepted Manuscript online:
February 06 2015
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2015 Biochemical Society
2015
Biochem J (2015) 467 (2): 333–343.
Article history
Received:
September 30 2014
Revision Received:
January 27 2015
Accepted:
February 06 2015
Accepted Manuscript online:
February 06 2015
Citation
Srividya Ganapathy, Odette Bécheau, Hanka Venselaar, Siebren Frölich, Jeroen B. van der Steen, Que Chen, Sarah Radwan, Johan Lugtenburg, Klaas J. Hellingwerf, Huub J. M. de Groot, Willem J. de Grip; Modulation of spectral properties and pump activity of proteorhodopsins by retinal analogues. Biochem J 15 April 2015; 467 (2): 333–343. doi: https://doi.org/10.1042/BJ20141210
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