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  1. Article ; Online: Heliorhodopsin binds and regulates glutamine synthetase activity.

    Cho, Shin-Gyu / Song, Myungchul / Chuon, Kimleng / Shim, Jin-Gon / Meas, Seanghun / Jung, Kwang-Hwan

    PLoS biology

    2022  Volume 20, Issue 10, Page(s) e3001817

    Abstract: Photoreceptors are light-sensitive proteins found in various organisms that respond to light and relay signals into the cells. Heliorhodopsin, a retinal-binding membrane protein, has been recently discovered, however its function remains unknown. Herein, ...

    Abstract Photoreceptors are light-sensitive proteins found in various organisms that respond to light and relay signals into the cells. Heliorhodopsin, a retinal-binding membrane protein, has been recently discovered, however its function remains unknown. Herein, we investigated the relationship between Actinobacteria bacterium IMCC26103 heliorhodopsin (AbHeR) and an adjacent glutamine synthetase (AbGS) in the same operon. We demonstrate that AbHeR binds to AbGS and regulates AbGS activity. More specifically, the dissociation constant (Kd) value of the binding between AbHeR and AbGS is 6.06 μM. Moreover, the absence of positively charged residues within the intracellular loop of AbHeR impacted Kd value as they serve as critical binding sites for AbGS. We also confirm that AbHeR up-regulates the biosynthetic enzyme activity of AbGS both in vitro and in vivo in the presence of light. GS is a key enzyme involved in nitrogen assimilation that catalyzes the conversion of glutamate and ammonia to glutamine. Hence, the interaction between AbHeR and AbGS may be critical for nitrogen assimilation in Actinobacteria bacterium IMCC26103 as it survives in low-nutrient environments. Overall, the findings of our study describe, for the first time, to the best of our knowledge, a novel function of heliorhodopsin as a regulatory rhodopsin with the capacity to bind and regulate enzyme activity required for nitrogen assimilation.
    MeSH term(s) Ammonia/metabolism ; Glutamate-Ammonia Ligase/chemistry ; Glutamate-Ammonia Ligase/metabolism ; Glutamic Acid/metabolism ; Glutamine ; Nitrogen ; Rhodopsin ; Rhodopsins, Microbial
    Chemical Substances Rhodopsins, Microbial ; heliorhodopsin ; Glutamine (0RH81L854J) ; Glutamic Acid (3KX376GY7L) ; Ammonia (7664-41-7) ; Rhodopsin (9009-81-8) ; Glutamate-Ammonia Ligase (EC 6.3.1.2) ; Nitrogen (N762921K75)
    Language English
    Publishing date 2022-10-03
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2126776-5
    ISSN 1545-7885 ; 1544-9173
    ISSN (online) 1545-7885
    ISSN 1544-9173
    DOI 10.1371/journal.pbio.3001817
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  2. Article ; Online: Mutational analyses identify a single amino acid critical for colour tuning in proteorhodopsins.

    Shim, Jin-Gon / Kang, Na-Rae / Chuon, Kimleng / Cho, Shin-Gyu / Meas, Seanghun / Jung, Kwang-Hwan

    FEBS letters

    2022  Volume 596, Issue 6, Page(s) 784–795

    Abstract: Microbial rhodopsins are light-activated proteins that contain seven transmembrane alpha-helices. Spectral tuning in microbial rhodopsins is a useful optogenetic tool. In this study, we report a new site that controls spectral tuning. In the ... ...

    Abstract Microbial rhodopsins are light-activated proteins that contain seven transmembrane alpha-helices. Spectral tuning in microbial rhodopsins is a useful optogenetic tool. In this study, we report a new site that controls spectral tuning. In the proteorhodopsins ISR34 and ISR36, a single amino-acid substitution at Cys189 caused an absorption maximum shift of 44 nm, indicating spectral tuning at a specific site. Comparison of single amino acid substitutions was conducted using photochemical and photobiological approaches. The maximum absorption for red-shift was measured for mutations at positions 189 and 105 in ISR34, both residues being equally important. Structural changes resulting from amino acid substitutions are related to pK
    MeSH term(s) Amino Acid Sequence ; Amino Acids/genetics ; Color ; Rhodopsin/chemistry ; Rhodopsins, Microbial/metabolism
    Chemical Substances Amino Acids ; Rhodopsins, Microbial ; proteorhodopsin ; Rhodopsin (9009-81-8)
    Language English
    Publishing date 2022-02-07
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 212746-5
    ISSN 1873-3468 ; 0014-5793
    ISSN (online) 1873-3468
    ISSN 0014-5793
    DOI 10.1002/1873-3468.14297
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  3. Article: Mutational analyses identify a single amino acid critical for colour tuning in proteorhodopsins

    Shim, Jin‐gon / Kang, Na‐rae / Chuon, Kimleng / Cho, Shin‐Gyu / Meas, Seanghun / Jung, Kwang‐Hwan

    FEBS letters. 2022 Mar., v. 596, no. 6

    2022  

    Abstract: Microbial rhodopsins are light‐activated proteins that contain seven transmembrane alpha‐helices. Spectral tuning in microbial rhodopsins is a useful optogenetic tool. In this study, we report a new site that controls spectral tuning. In the ... ...

    Abstract Microbial rhodopsins are light‐activated proteins that contain seven transmembrane alpha‐helices. Spectral tuning in microbial rhodopsins is a useful optogenetic tool. In this study, we report a new site that controls spectral tuning. In the proteorhodopsins ISR34 and ISR36, a single amino‐acid substitution at Cys189 caused an absorption maximum shift of 44 nm, indicating spectral tuning at a specific site. Comparison of single amino acid substitutions was conducted using photochemical and photobiological approaches. The maximum absorption for red‐shift was measured for mutations at positions 189 and 105 in ISR34, both residues being equally important. Structural changes resulting from amino acid substitutions are related to pKₐ values, pumping activity and spectral tuning.
    Keywords absorption ; amino acids ; color ; optogenetics ; photochemistry ; rhodopsin
    Language English
    Dates of publication 2022-03
    Size p. 784-795.
    Publishing place John Wiley & Sons, Ltd
    Document type Article
    Note JOURNAL ARTICLE
    ZDB-ID 212746-5
    ISSN 1873-3468 ; 0014-5793
    ISSN (online) 1873-3468
    ISSN 0014-5793
    DOI 10.1002/1873-3468.14297
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  4. Article ; Online: Heliorhodopsin Helps Photolyase to Enhance the DNA Repair Capacity.

    Shim, Jin-Gon / Cho, Shin-Gyu / Kim, Se-Hwan / Chuon, Kimleng / Meas, Seanghun / Choi, Ahreum / Jung, Kwang-Hwan

    Microbiology spectrum

    2022  Volume 10, Issue 6, Page(s) e0221522

    Abstract: Light quality is a significant factor for living organisms that have photosensory systems, such as rhodopsin, a seven alpha-helical transmembrane protein with the retinal chromophore. Here, we report, for the first time, the function of new rhodopsin, ... ...

    Abstract Light quality is a significant factor for living organisms that have photosensory systems, such as rhodopsin, a seven alpha-helical transmembrane protein with the retinal chromophore. Here, we report, for the first time, the function of new rhodopsin, which is an inverted 7-transmembrane protein, isolated from Trichococcus flocculiformis
    MeSH term(s) Deoxyribodipyrimidine Photo-Lyase/chemistry ; Deoxyribodipyrimidine Photo-Lyase/genetics ; Deoxyribodipyrimidine Photo-Lyase/metabolism ; Rhodopsin/genetics ; Pyrimidine Dimers/chemistry ; Pyrimidine Dimers/metabolism ; DNA Repair
    Chemical Substances Deoxyribodipyrimidine Photo-Lyase (EC 4.1.99.3) ; heliorhodopsin ; Rhodopsin (9009-81-8) ; Pyrimidine Dimers
    Language English
    Publishing date 2022-10-11
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2807133-5
    ISSN 2165-0497 ; 2165-0497
    ISSN (online) 2165-0497
    ISSN 2165-0497
    DOI 10.1128/spectrum.02215-22
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  5. Article ; Online: Color-tuning of natural variants of heliorhodopsin.

    Kim, Se-Hwan / Chuon, Kimleng / Cho, Shin-Gyu / Choi, Ahreum / Meas, Seanghun / Cho, Hyun-Suk / Jung, Kwang-Hwan

    Scientific reports

    2021  Volume 11, Issue 1, Page(s) 854

    Abstract: Microbial rhodopsins are distributed through many microorganisms. Heliorhodopsins are newly discovered but have an unclear function. They have seven transmembrane helices similar to type-I and type-II rhodopsins, but they are different in that the N- ... ...

    Abstract Microbial rhodopsins are distributed through many microorganisms. Heliorhodopsins are newly discovered but have an unclear function. They have seven transmembrane helices similar to type-I and type-II rhodopsins, but they are different in that the N-terminal region of heliorhodopsin is cytoplasmic. We chose 13 representative heliorhodopsins from various microorganisms, expressed and purified with an N-terminal His tag, and measured the absorption spectra. The 13 natural variants had an absorption maximum (λmax) in the range 530-556 nm similar to proteorhodopsin (λmax = 490-525 nm). We selected several candidate residues that influence rhodopsin color-tuning based on sequence alignment and constructed mutants via site-directed mutagenesis to confirm the spectral changes. We found two important residues located near retinal chromophore that influence λmax. We also predict the 3D structure via homology-modeling of Thermoplasmatales heliorhodopsin. The results indicate that the color-tuning mechanism of type-I rhodopsin can be applied to understand the color-tuning of heliorhodopsin.
    Language English
    Publishing date 2021-01-13
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-020-72125-0
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  6. Article ; Online: Discovery of a new light-driven Li

    Cho, Shin-Gyu / Shim, Jin-Gon / Choun, Kimleng / Meas, Seanghun / Kang, Kun-Wook / Kim, Ji-Hyun / Cho, Hyun-Suk / Jung, Kwang-Hwan

    Journal of photochemistry and photobiology. B, Biology

    2021  Volume 223, Page(s) 112285

    Abstract: Microbial pumping rhodopsin is a seven-transmembrane retinal binding protein, which is light-driven ion pump with a functional key motif. Ion-pumping with the key motif and charged amino acids in the rhodopsin is biochemically important. The rhodopsins ... ...

    Abstract Microbial pumping rhodopsin is a seven-transmembrane retinal binding protein, which is light-driven ion pump with a functional key motif. Ion-pumping with the key motif and charged amino acids in the rhodopsin is biochemically important. The rhodopsins with DTG motif have been discovered in various eubacteria, and they function as H
    MeSH term(s) Amino Acid Motifs ; Hydrogen-Ion Concentration ; Ion Transport/radiation effects ; Light ; Lithium/chemistry ; Lithium/metabolism ; Methylobacteriaceae/metabolism ; Mutagenesis, Site-Directed ; Phylogeny ; Protein Binding ; Protein Conformation, alpha-Helical ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/chemistry ; Recombinant Proteins/isolation & purification ; Rhodopsins, Microbial/chemistry ; Rhodopsins, Microbial/classification ; Rhodopsins, Microbial/genetics ; Rhodopsins, Microbial/metabolism ; Sodium/chemistry ; Sodium/metabolism
    Chemical Substances Recombinant Proteins ; Rhodopsins, Microbial ; Lithium (9FN79X2M3F) ; Sodium (9NEZ333N27)
    Language English
    Publishing date 2021-08-12
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 623022-2
    ISSN 1873-2682 ; 1011-1344
    ISSN (online) 1873-2682
    ISSN 1011-1344
    DOI 10.1016/j.jphotobiol.2021.112285
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  7. Article: Discovery of a new light-driven Li+/Na+-pumping rhodopsin with DTG motif

    Cho, Shin-Gyu / Shim, Jin-gon / Choun, Kimleng / Meas, Seanghun / Kang, Kun-Wook / Kim, Ji-hyun / Cho, Hyun-Suk / Jung, Kwang-Hwan

    Journal of photochemistry and photobiology. 2021 Oct., v. 223

    2021  

    Abstract: Microbial pumping rhodopsin is a seven-transmembrane retinal binding protein, which is light-driven ion pump with a functional key motif. Ion-pumping with the key motif and charged amino acids in the rhodopsin is biochemically important. The rhodopsins ... ...

    Abstract Microbial pumping rhodopsin is a seven-transmembrane retinal binding protein, which is light-driven ion pump with a functional key motif. Ion-pumping with the key motif and charged amino acids in the rhodopsin is biochemically important. The rhodopsins with DTG motif have been discovered in various eubacteria, and they function as H⁺ pump. Especially, the DTG motif rhodopsins transported H⁺ despite the replacement of a proton donor by Gly. We investigated Methylobacterium populi rhodopsin (MpR) in one of the DTG motif rhodopsin clades. To determine which ions the MpR transport, we tested with various monovalent ion solutions and determined that MpR transports Li⁺/Na⁺. By replacing the three negatively charged residues residues which are located in helix B, Glu32, Glu33, and Asp35, we concluded that the residues play a critical role in the transport of Li⁺/Na⁺. The MpR E33Q transported H⁺ in place of Li⁺/Na⁺, suggesting that Glu33 is a Li⁺/Na⁺ binding site on the cytoplasmic side. Gly93 in MpR was replaced by Asp to convert from the Li⁺/Na⁺ pump to the H⁺ pump, resulting in MpR G93D transporting H⁺. Dissociation constant (Kd) values of Na⁺ for MpR WT and E33Q were determined to be 4.0 and 72.5 mM, respectively. These results indicated the mechanism by which MpR E33Q transports H⁺. Up to now, various ion-pumping rhodopsins have been discovered, and Li⁺/Na⁺-pumping rhodopsins were only found in the NDQ motif in NaR. Here, we report a new light-driven Na⁺ pump MpR and have determined the important residues required for Li⁺/Na⁺-pumping different from previously known NaR.
    Keywords Methylobacterium ; photobiology ; photochemistry ; rhodopsin
    Language English
    Dates of publication 2021-10
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 623022-2
    ISSN 1873-2682 ; 1011-1344
    ISSN (online) 1873-2682
    ISSN 1011-1344
    DOI 10.1016/j.jphotobiol.2021.112285
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  8. Article: The role of carotenoids in proton-pumping rhodopsin as a primitive solar energy conversion system

    Chuon, Kimleng / Shim, Jin-gon / Kim, Se-Hwan / Cho, Shin-Gyu / Meas, Seanghun / Kang, Kun-Wook / Kim, Ji-Hyun / Das, Ishita / Sheves, Mordechai / Jung, Kwang-Hwan

    Journal of photochemistry and photobiology. 2021 Aug., v. 221

    2021  

    Abstract: Rhodopsin and carotenoids are two molecules that certain bacteria use to absorb and utilize light. Type I rhodopsin, the simplest active proton transporter, converts light energy into an electrochemical potential. Light produces a proton gradient, which ... ...

    Abstract Rhodopsin and carotenoids are two molecules that certain bacteria use to absorb and utilize light. Type I rhodopsin, the simplest active proton transporter, converts light energy into an electrochemical potential. Light produces a proton gradient, which is known as the proton motive force across the cell membrane. Some carotenoids are involved in light absorbance and transfer of absorbed energy to chlorophyll during photosynthesis. A previous study in Salinibacter ruber has shown that carotenoids act as antennae to harvest light and transfer energy to retinal in xanthorhodopsin (XR). Here, we describe the role of canthaxanthin (CAN), a carotenoid, as an antenna for Gloeobacter rhodopsin (GR). The non-covalent complex formed by the interaction between CAN and GR doubled the proton pumping speed and improved the pumping capacity by 1.5-fold. The complex also tripled the proton pumping speed and improved the pumping capacity by 5-fold in the presence of strong and weak light, respectively. Interestingly, when canthaxanthin was bound to Gloeobacter rhodopsin, it showed a 126-fold increase in heat resistance, and it survived better under drought conditions than Gloeobacter rhodopsin. The results suggest direct complementation of Gloeobacter rhodopsin with a carotenoid for primitive solar energy harvesting in cyanobacteria.
    Keywords Gloeobacter ; Salinibacter ruber ; absorbance ; canthaxanthin ; cell membranes ; chlorophyll ; drought ; electrochemistry ; energy conversion ; heat tolerance ; photobiology ; photochemistry ; photosynthesis ; proton-motive force ; rhodopsin ; solar energy
    Language English
    Dates of publication 2021-08
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 623022-2
    ISSN 1873-2682 ; 1011-1344
    ISSN (online) 1873-2682
    ISSN 1011-1344
    DOI 10.1016/j.jphotobiol.2021.112241
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  9. Article ; Online: Carotenoid binding in Gloeobacteria rhodopsin provides insights into divergent evolution of xanthorhodopsin types.

    Chuon, Kimleng / Shim, Jin-Gon / Kang, Kun-Wook / Cho, Shin-Gyu / Hour, Chenda / Meas, Seanghun / Kim, Ji-Hyun / Choi, Ahreum / Jung, Kwang-Hwan

    Communications biology

    2022  Volume 5, Issue 1, Page(s) 512

    Abstract: The position of carotenoid in xanthorhodopsin has been elucidated. However, a challenging expression of this opsin and a complex biosynthesis carotenoid in the laboratory hold back the insightful study of this rhodopsin. Here, we demonstrated co- ... ...

    Abstract The position of carotenoid in xanthorhodopsin has been elucidated. However, a challenging expression of this opsin and a complex biosynthesis carotenoid in the laboratory hold back the insightful study of this rhodopsin. Here, we demonstrated co-expression of the xanthorhodopsin type isolated from Gloeobacter violaceus PCC 7421-Gloeobacter rhodopsin (GR) with a biosynthesized keto-carotenoid (canthaxanthin) targeting the carotenoid binding site. Direct mutation-induced changes in carotenoid-rhodopsin interaction revealed three crucial features: (1) carotenoid locked motif (CLM), (2) carotenoid aligned motif (CAM), and color tuning serines (CTS). Our single mutation results at 178 position (G178W) confirmed inhibition of carotenoid binding; however, the mutants showed better stability and proton pumping, which was also observed in the case of carotenoid binding characteristics. These effects demonstrated an adaptation of microbial rhodopsin that diverges from carotenoid harboring, along with expression in the dinoflagellate Pyrocystis lunula rhodopsin and the evolutionary substitution model. The study highlights a critical position of the carotenoid binding site, which significantly allows another protein engineering approach in the microbial rhodopsin family.
    MeSH term(s) Binding Sites ; Carotenoids/metabolism ; Proton Pumps ; Rhodopsin/genetics ; Rhodopsin/metabolism ; Rhodopsins, Microbial/genetics ; Rhodopsins, Microbial/metabolism
    Chemical Substances Proton Pumps ; Rhodopsins, Microbial ; Carotenoids (36-88-4) ; Rhodopsin (9009-81-8)
    Language English
    Publishing date 2022-05-30
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2399-3642
    ISSN (online) 2399-3642
    DOI 10.1038/s42003-022-03429-2
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  10. Article: Assembly of Natively Synthesized Dual Chromophores Into Functional Actinorhodopsin.

    Chuon, Kimleng / Kim, So Young / Meas, Seanghun / Shim, Jin-Gon / Cho, Shin-Gyu / Kang, Kun-Wook / Kim, Ji-Hyun / Cho, Hyun-Suk / Jung, Kwang-Hwan

    Frontiers in microbiology

    2021  Volume 12, Page(s) 652328

    Abstract: Microbial rhodopsin is a simple solar energy-capturing molecule compared to the complex photosynthesis apparatus. Light-driven proton pumping across the cell membrane is a crucial mechanism underlying microbial energy production. ...

    Abstract Microbial rhodopsin is a simple solar energy-capturing molecule compared to the complex photosynthesis apparatus. Light-driven proton pumping across the cell membrane is a crucial mechanism underlying microbial energy production.
    Language English
    Publishing date 2021-04-28
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2587354-4
    ISSN 1664-302X
    ISSN 1664-302X
    DOI 10.3389/fmicb.2021.652328
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