Modulation of spectral properties and pump activity of proteorhodopsins by retinal analogues

Ganapathy, Srividya; Bécheau, Odette; Venselaar, Hanka; Frölich, Siebren; Steen, Jeroen B. van der; Chen, Que; Radwan, Sarah; Lugtenburg, J.; Hellingwerf, Klaas J.; Groot, Huub J. M. de; Grip, W.J. de

doi:10.1042/bj20141210

Cited by 27 publications

(64 citation statements)

References 63 publications

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“…The first is to use a proteorhodopsin variant that pumps faster. Since the start of this project several routes towards improvement along this line have revealed themselves, like the use of proteorhodopsin variants that bind a carotenoid like echinenone (84), not only because of their pumping rate (32), but also because their antenna carotenoid increases their effective absorption cross section 3.5 fold (85). A second possibility may be to use the trimeric bacteriorhodopsin, which presumably will not form an extended hexagonal lattice, but will show a higher pumping rate.…”

Section: Discussionmentioning

confidence: 99%

“…For this, the absorbance maximum of the proteorhodopsin would have to be shifted to beyond 700 nm. Although progress is made in this direction (32) there is a significant way still to go. In a seminal paper, written at the peak of the recent revival of interest in sustainability research (33), a comparison of 'artificial' and natural photosynthesis was made, together with a review of options to increase the efficiency of light energy conversion through rational engineering in both approaches.…”

Section: Discussionmentioning

confidence: 99%

“…Introduction of PRs into such a system could lead to increased CO 2 fixation, which could ultimately increase production rates of interesting compounds by cyanobacterial cell factories (31). Hence, it has previously been suggested that PRs could supply additional free energy to oxyphototrophic organisms (29,30), even more so when it turns out to be possible to shift the window of absorption of this pigmented protein outside that of the PAR region (32). Several proposals have been made to increase the efficiency of oxygenic photosynthesis beyond its natural limits (e.g.…”

Section: Introductionmentioning

confidence: 99%

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Expression of holo-proteorhodopsin in Synechocystis sp. PCC 6803

Chen

Steen

Dekker

et al. 2016

Metabolic Engineering

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Retinal-based photosynthesis may contribute to the free energy conversion needed for growth of an organism carrying out oxygenic photosynthesis, like a cyanobacterium. After optimization, this may even enhance the overall efficiency of phototrophic growth of such organisms in sustainability applications. As a first step towards this, we here report on functional expression of the archetype proteorhodopsin in Synechocystis sp. PCC 6803. Upon use of the moderate-strength psbA2 promoter, holo-proteorhodopsin is expressed in this cyanobacterium, at a level of up to 10 5 molecules per cell, presumably in a hexameric quaternary structure, and with approximately equal distribution (on a protein-content basis) over the thylakoid and the cytoplasmic membrane fraction. These results also demonstrate that Synechocystis sp. PCC 6803 has the capacity to synthesize alltrans-retinal. Expressing a substantial amount of a heterologous opsin membrane protein causes a substantial growth retardation Synechocystis, as is clear from a strain expressing PROPS, a nonpumping mutant derivative of proteorhodopsin. Relative to this latter strain, proteorhodopsin expression, however, measurably stimulates its growth.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression of holo-proteorhodopsin in Synechocystis sp. PCC 6803

Chen

Steen

Dekker

et al. 2016

Metabolic Engineering

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“…We have previously demonstrated red and blue shifts in the absorbance bands of PR, GR, and PR-DNFS using analogues of A1 with different ring modifications. 29 Of these, the analogue all- trans -3,4-dehydroretinal (A2) induced a significant red shift (26–32 nm) in all pigments while largely retaining the pump activity. 29 A2 is the only other naturally occurring retinal, and it has been found in the photoreceptor cells of certain fish, invertebrates, and amphibians.…”

Section: Introductionmentioning

confidence: 99%

Retinal-Based Proton Pumping in the Near Infrared

Venselaar²,

et al. 2017

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Proteorhodopsin (PR) and Gloeobacter rhodopsin (GR) are retinal-based light-driven proton pumps that absorb visible light (maxima at 520–540 nm). Shifting the action spectra of these proton pumps beyond 700 nm would generate new prospects in optogenetics, membrane sensor technology, and complementation of oxygenic phototrophy. We therefore investigated the effect of red-shifting analogues of retinal, combined with red-shifting mutations, on the spectral properties and pump activity of the resulting pigments. We investigated a variety of analogues, including many novel ones. One of the novel analogues we tested, 3-methylamino-16-nor-1,2,3,4-didehydroretinal (MMAR), produced exciting results. This analogue red-shifted all of the rhodopsin variants tested, accompanied by a strong broadening of the absorbance band, tailing out to 850–950 nm. In particular, MMAR showed a strong synergistic effect with the PR-D212N,F234S double mutant, inducing an astonishing 200 nm red shift in the absorbance maximum. To our knowledge, this is by far the largest red shift reported for any retinal protein. Very importantly, all MMAR-containing holoproteins are the first rhodopsins retaining significant pump activity under near-infrared illumination (730 nm light-emitting diode). Such MMAR-based rhodopsin variants present very promising opportunities for further synthetic biology modification and for a variety of biotechnological and biophysical applications.

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“…Toward this purpose, two types of red-shifted ChRs have been made: (i) mutation of amino acids in the vicinity of the retinal pocket, [9][10][11] and (ii) substitution of retinal with other red-shifted chromophores that covalently bind to ChRs through a PSB. [12][13][14] In this Communication, we present new types of redshifted ChRs: red-shifted chromophores 3 and 4, consisting of enamine-conjugated Schiff bases of 1 and 3,4-dehydroretinal (2), that non-covalently binds to chimera ChRs (Fig. 1b).…”

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confidence: 99%

Alternative Formation of Red-Shifted Channelrhodopsins: Noncovalent Incorporation with Retinal-Based Enamine-Type Schiff Bases and Mutated Channelopsin

Okitsu

Matsuyama

Yamashita

et al. 2017

CHEMICAL & PHARMACEUTICAL BULLETIN

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Red-shifted channelrhodopsins (ChRs) are attractive for optogenetic tools. We developed a new type of red-shifted ChRs that utilized noncovalent incorporation of retinal and 3,4-dehydroretinal-based enamine-type Schiff bases and mutated channelopsin, C1C2-K296G. These ChRs exhibited absorption maxima that were shifted 10-30 nm toward longer wavelengths than that of C1C2-ChR regenerated with alltrans-retinal.Key words channelrhodopsin; chromophore; red-shift; neuroscience; optogenetics The development of a new method of activating a specific neuron without affecting other cells would greatly benefit neuroscience. Optogenetics-a powerful new technique that enables control of neuronal activity by light-offers a number of advantages over previous electrode-based methods.1-4) In addition, optogenetics appears promising for clarifying neural circuits and understanding the mechanisms of various neurological diseases. 5,6) Channelrhodopsins (ChRs) are members of the rhodopsin family in microbes and consist of the chromophore retinal (1) and a seven-transmembrane helix apoprotein, termed channelopsin (Chop). The amino group of a lysine residue (K296) covalently binds to retinal through a protonated Schiff base (PSB) 7) (Fig. 1a). ChRs function as light-gated cation channels by photoisomerization of the retinal from all-trans to 13-cis forms which results in a conformational change of the protein. Two ChRs, Channelrhodopsin-1 (ChR1) and Channelrhodopsin-2 (ChR2), are now widely used in optogenetics, and they are most sensitive to green and blue light, respectively. 7,8) However, such short-wavelength light cannot deeply penetrate into the neural tissues, and optical fiber implants are often required to target the tissues. A truly noninvasive method for optogenetics requires the development of ChRs that are sensitive to long-wavelength light, and so red-shifted ChRs are sought-after. Toward this purpose, two types of red-shifted ChRs have been made: (i) mutation of amino acids in the vicinity of the retinal pocket, [9][10][11] and (ii) substitution of retinal with other red-shifted chromophores that covalently bind to ChRs through a PSB. [12][13][14] In this Communication, we present new types of redshifted ChRs: red-shifted chromophores 3 and 4, consisting of enamine-conjugated Schiff bases of 1 and 3,4-dehydroretinal (2), that non-covalently binds to chimera ChRs (Fig. 1b). This new red-shifted ChR system would have two benefits: (i) introduction of one or two conjugated double bond(s) into the chromophore would result in absorption of longer wavelength light for a red-shift of the UV-Vis spectrum, and (ii) instead of a covalent bond, formation of the selective chromophorechimera Chop would occur by action of van der Waals forces.Herein, we designed chromophores 3 and 4, which we expected to have red-shifted absorption maxima compared to 1 because the presence of the additional double bonds extended the conjugation of the chromophore.15) The most difficult problems for this study were (i) construction of the enamine-conj...

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Modulation of spectral properties and pump activity of proteorhodopsins by retinal analogues

Cited by 27 publications

References 63 publications

Expression of holo-proteorhodopsin in Synechocystis sp. PCC 6803

Expression of holo-proteorhodopsin in Synechocystis sp. PCC 6803

Retinal-Based Proton Pumping in the Near Infrared

Alternative Formation of Red-Shifted Channelrhodopsins: Noncovalent Incorporation with Retinal-Based Enamine-Type Schiff Bases and Mutated Channelopsin

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