Abstract
The UV light absorbing species of proteorhodopsin with deprotonated Schiff base retinal was investigated using steady-state fluorescence and femtosecond pump-probe spectroscopy. Compared to the all-trans retinal with protonated Schiff base, the deprotonated chromophore absorbs at 365 nm and exhibits a blue-shifted fluorescence spectrum. The unusually long-lived excited state decays bi-exponentially with time constants of 8 ps and 130 ps to form a deprotonated 13-cis retinal as the primary photo-product.
Acknowledgments
This work has been funded by the DFG via SFB 807 ‘Transport and Communication across Biological Membranes’. We thank Maria Dell for her contributions to the titration experiments.
References
Bamann, C., Bamberg, E., Wachtveitl, J., and Glaubitz, C. (2014). Proteorhodopsin. BBA-Bioenergetics 1837, 614–625.10.1016/j.bbabio.2013.09.010Search in Google Scholar
Béjà, O., Aravind, L., Koonin, E.V., Suzuki, M.T., Hadd, A., Nguyen, L.P., Jovanovich, S.B., Gates, C.M., Feldmann, R.A., Spudich, J.L., et al. (2000). Bacterial rhodopsin: evidence for a new type of phototrophy in the sea. Science 289, 1902–1906.10.1126/science.289.5486.1902Search in Google Scholar
Béjà, O., Spudich, E.N., Spudich, J.L., Leclerc, M., and DeLong, E.F. (2001). Proteorhodopsin phototrophy in the ocean. Nature 411, 786–789.10.1038/35081051Search in Google Scholar
de la Torre, J.R., Christianson, L.M., Béjà, O., Suzuki, M.T., Karl, D.M., Heidelberg, J., and DeLong, E.F. (2003). Proteorhodopsin genes are distributed among divergent marine bacterial taxa. Proc. Natl. Acad. Sci. USA 100, 12830–12835.10.1073/pnas.2133554100Search in Google Scholar
Friedrich, T., Geibel, S., Kalmbach, R., Chizhov, I., Ataka, K., Heberle, J., Engelhard, M., and Bamberg, E. (2002). Proteorhodopsin is a light-driven proton pump with variable vectoriality. J. Mol. Biol. 321, 821–838.10.1016/S0022-2836(02)00696-4Search in Google Scholar
Hempelmann, F., Hölper, S., Verhoefen, M.-K., Woerner, A.C., Köhler, T., Fiedler, S.-A., Pfleger, N., Wachtveitl, J., and Glaubitz, C. (2011). His75-Asp97 cluster in green proteorhodopsin. J. Am. Chem. Soc. 133, 4645–4654.10.1021/ja111116aSearch in Google Scholar PubMed
Herz, J., Verhoefen, M.-K., Weber, I., Bamann, C., Glaubitz, C., and Wachtveitl, J. (2012). Critical role of Asp227 in the photocycle of proteorhodopsin. Biochemistry 51, 5589–5600.10.1021/bi3003764Search in Google Scholar PubMed
Huber, R., Köhler, T., Lenz, M.O., Bamberg, E., Kalmbach, R., Engelhardt, M., and Wachtveitl, J. (2005). pH-Dependent photoisomerization of retinal in proteorhodopsin. Biochemistry 44, 1800–1806.10.1021/bi048318hSearch in Google Scholar PubMed
Imasheva, E.S., Balashov, S.P., Wang, J.M., Dioumaev, A.K., and Lanyi, J.K. (2004). Selectivity of Retinal photoisomerization in proteorhodopsin is controlled by aspartatic acid 227. Biochemistry 43, 1648–1655.10.1021/bi0355894Search in Google Scholar PubMed
Imasheva, E.S., Shimono, K., Balashov, S.P., Wang, J.M., Zadok, U., Sheves, M., Kamo, N., and Lanyi, K. (2005). Formation of a long-lived photoproduct with a deprotonated Schiff base in proteorhodopsin, and its enhancement by mutation of Asp227. Biochemistry 44, 10828–10838.10.1021/bi050438hSearch in Google Scholar PubMed
Kovalenko, S.A., Dobryakov, A.L., Ruthmann, J., and Ernsting, N.P. (1999). Femtosecond spectroscopy of condenced phases with chirped supercontinuum probing. Phys. Rev. A 59, 2369–2384.10.1103/PhysRevA.59.2369Search in Google Scholar
Kozma, I.Z., Baum, P., Lochbrunner, S., and Riedle, E. (2003). Widely tunable sub-30 fs ultraviolet pulses by chirped sum frequency mixing. Opt. Express 11, 3110–3115.10.1364/OE.11.003110Search in Google Scholar PubMed
Kralj, J.M., Spudich, E.N., Spudich, J.L., and Rothschild, K.J. (2008). Raman spectroscopy reveals direct chromophore interactions in the Leu/Gln105 spectral tuning switch of proteorhodopsins. J. Phys. Chem. B 112, 11770–11776.10.1021/jp802629eSearch in Google Scholar PubMed PubMed Central
Lenz, M.O., Huber, R., Schmidt, B., Gilch, P., Kalmbach, R., Engelhard, M., and Wachtveitl, J. (2006). First steps of retinal photoisomerization in proteorhodopsin. Biophys. J. 91, 255–262.10.1529/biophysj.105.074690Search in Google Scholar PubMed PubMed Central
Lörinczi, É., Verhoefen, M.-K., Wachtveitl, J., Woerner, A.C., Glaubitz, C., Engelhard, M., Bamberg, E., and Friedrich, T. (2009). Voltage- and pH-dependent changes in vectoriality of photocurrents mediated by wild-type and mutant proteorhodopsins upon expression in Xenopus oocytes. J. Mol. Biol. 393, 320–341.10.1016/j.jmb.2009.07.055Search in Google Scholar PubMed
Luck, M., Mathes, T., Bruun, S., Fudim, R., Hagedorn, R., Tran Nguyen, T.M., Kateriya, S., Kennis, J.T.M., Hildebrandt, P., and Hegemann, P. (2012). A photochromic histidine kinase rhodopsin (HKR1) that is bimodally switched by ultraviolet and blue light. J. Biol. Chem. 287, 40083–40090.10.1074/jbc.M112.401604Search in Google Scholar PubMed PubMed Central
Man, D., Wang, W., Sabehi, G., Aravind, L., Post, A.F., Massana, R., Spudich, E.N., Spudich, J.L., and Béjà, O. (2003). Diversification and spectral tuning in marine proteorhodopsins. EMBO J. 22, 1725–1731.10.1093/emboj/cdg183Search in Google Scholar PubMed PubMed Central
Mao, J., Do, N.-N., Scholz, F., Reggie, L., Mehler, M., Lakatos, A., Ong, Y.-S., Ullrich, S.J., Brown, L.J., Brown, R.C.D., et al. (2014). Structural basis of the green-blue color switching in proteorhodopsin as determined by NMR spectroskopy. J. Am. Chem. Soc. 136, 17578–17590.10.1021/ja5097946Search in Google Scholar PubMed
Nielsen, I.B., Lammich, L., and Andersen, L.H. (2006). S1 and S2 excited states of gas-phase Schiff-base retinal chromophores. Phys. Rev. Lett. 96, Article no. 018304, 1–4.Search in Google Scholar
Pfleger, N., Lorch, M., Woerner, A.C., Shastri, S., and Glaubitz, C. (2008). Characterization of Schiff base and chromophore in green proteorhodopsin by solid-state NMR. J. Biomol. NMR 40, 15–21.10.1007/s10858-007-9203-5Search in Google Scholar PubMed
Riedle, E., Beutter, M., Lochbrunner, S., Piel, J., Schenkl, S., Spörlein, S., and Zinth, W. (2000). Generation of 10 to 50 fs pulses tunable through all of the visible and the NIR. Appl. Phys. B 71, 457–465.10.1007/s003400000351Search in Google Scholar
Slavov, C., Hartmann, H., and Wachtveitl, J. (2015). Implementation and evaluation of data analysis strategies for time-resolved optical spectroscopy. Analyt. Chem. 87, 2328–2336.10.1021/ac504348hSearch in Google Scholar PubMed
van Stokkum, I.H., Larsen, D.S., and van Grondelle, R. (2004). Global and target analysis of time-resolved spectra. Biochim. Biophys. Acta 1657, 82–104.10.1016/j.bbabio.2004.04.011Search in Google Scholar PubMed
Verhoefen, M.-K., Schäfer, G., Shastri, S., Weber, I., Glaubitz, C., Mäntele, W., and Wachtveitl, J. (2011). Low temperature FTIR spectroscopy provides new insights in the pH-dependent proton pathway of proteorhodopsin. BBA Bioenergetics 1807, 1583–1590.10.1016/j.bbabio.2011.09.001Search in Google Scholar PubMed
Yang, J., Aslimovska, L., and Glaubitz, C. (2011). Molecular dynamics of proteorhodopsin in lipid bilayers by solid-state NMR. J. Am. Chem. Soc. 133, 4874–4881.10.1021/ja109766nSearch in Google Scholar PubMed
Supplemental Material:
The online version of this article (DOI: 10.1515/hsz-2015-0108) offers supplementary material, available to authorized users.
©2015 by De Gruyter
Articles in the same Issue
- Frontmatter
- Meeting Report
- Membrane Transport and Communication in Frankfurt: Speakers’ Summary – Highlights
- HIGHLIGHT: MEMBRANE TRANSPORT AND COMMUNICATION
- Structure, function, evolution, and application of bacterial Pnu-type vitamin transporters
- Team work at its best – TAPL and its two domains
- The volume-regulated anion channel is formed by LRRC8 heteromers – molecular identification and roles in membrane transport and physiology
- Extending native mass spectrometry approaches to integral membrane proteins
- Functional diversity of the superfamily of K+ transporters to meet various requirements
- The structure of Na+-translocating of NADH:ubiquinone oxidoreductase of Vibrio cholerae: implications on coupling between electron transfer and Na+ transport
- Hybrid rotors in F1Fo ATP synthases: subunit composition, distribution, and physiological significance
- Homeostatic control of biological membranes by dedicated lipid and membrane packing sensors
- The transporter associated with antigen processing: a key player in adaptive immunity
- The pseudo-atomic structure of an RND-type tripartite multidrug efflux pump
- The assembly and disassembly of the AcrAB-TolC three-component multidrug efflux pump
- A universal mechanism for transport and regulation of CPA sodium proton exchangers
- Biosynthesis of membrane dependent proteins in insect cell lysates: identification of limiting parameters for folding and processing
- Fluorescence and excited state dynamics of the deprotonated Schiff base retinal in proteorhodopsin
- Regulatory role of charged clusters in the N-terminal domain of BetP from Corynebacterium glutamicum
- The contribution of methionine to the stability of the Escherichia coli MetNIQ ABC transporter-substrate binding protein complex
- The ABC exporter MsbA probed by solid state NMR – challenges and opportunities
- Functional properties of LptA and LptD in Anabaena sp. PCC 7120
Articles in the same Issue
- Frontmatter
- Meeting Report
- Membrane Transport and Communication in Frankfurt: Speakers’ Summary – Highlights
- HIGHLIGHT: MEMBRANE TRANSPORT AND COMMUNICATION
- Structure, function, evolution, and application of bacterial Pnu-type vitamin transporters
- Team work at its best – TAPL and its two domains
- The volume-regulated anion channel is formed by LRRC8 heteromers – molecular identification and roles in membrane transport and physiology
- Extending native mass spectrometry approaches to integral membrane proteins
- Functional diversity of the superfamily of K+ transporters to meet various requirements
- The structure of Na+-translocating of NADH:ubiquinone oxidoreductase of Vibrio cholerae: implications on coupling between electron transfer and Na+ transport
- Hybrid rotors in F1Fo ATP synthases: subunit composition, distribution, and physiological significance
- Homeostatic control of biological membranes by dedicated lipid and membrane packing sensors
- The transporter associated with antigen processing: a key player in adaptive immunity
- The pseudo-atomic structure of an RND-type tripartite multidrug efflux pump
- The assembly and disassembly of the AcrAB-TolC three-component multidrug efflux pump
- A universal mechanism for transport and regulation of CPA sodium proton exchangers
- Biosynthesis of membrane dependent proteins in insect cell lysates: identification of limiting parameters for folding and processing
- Fluorescence and excited state dynamics of the deprotonated Schiff base retinal in proteorhodopsin
- Regulatory role of charged clusters in the N-terminal domain of BetP from Corynebacterium glutamicum
- The contribution of methionine to the stability of the Escherichia coli MetNIQ ABC transporter-substrate binding protein complex
- The ABC exporter MsbA probed by solid state NMR – challenges and opportunities
- Functional properties of LptA and LptD in Anabaena sp. PCC 7120