Skip to main content

Advertisement

SpringerLink
Log in

Exploring color tuning strategies in red fluorescent proteins

  • Perspective
  • Published:
Photochemical & Photobiological Sciences Aims and scope Submit manuscript

Abstract

Red-emitting fluorescent proteins (RFPs) with fluorescence emission above 600 nm are advantageous for cell and tissue imaging applications for various reasons. Fluorescence from an RFP is well separated from cellular autofluorescence, which is in the green region of the spectrum, and red light is scattered less, which allows thicker specimens to be imaged. Moreover, the phototoxic response of cells is lower for red than blue or green light exposure. Further red-shifted FP variants can be obtained by genetic modifications causing an extension of the conjugated p-electron system of the chromophore, or by placing amino acids near the chromophore that stabilize its excited state or destabilize its ground state. We have selected the tetrameric RFP eqFP611 from Entacmaea quadricolor as a lead structure and discuss several rational design trials to generate RFP variants with improved photochemical properties.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. O. Shimomura, F. H. Johnson and Y. Saiga, Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea J. Cell. Comp. Physiol. 1962 59 223.

    Article  CAS  PubMed  Google Scholar 

  2. O. Shimomura, Structure of the Chromophore of Aequorea Green Fluorescent Protein FEBS Lett. 1979 104 220.

    Article  CAS  Google Scholar 

  3. D. C. Prasher, V. K. Eckenrode, W. W. Ward, F. G. Prendergast and M. J. Cormier, Primary structure of the Aequorea victoria green-fluorescent protein Gene 1992 111 229.

    Article  CAS  PubMed  Google Scholar 

  4. M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward and D. C. Prasher, Green fluorescent protein as a marker for gene expression Science 1994 263 802.

    Article  CAS  PubMed  Google Scholar 

  5. R. Y. Tsien, The green fluorescent protein Annu. Rev. Biochem. 1998 67 509.

    Article  CAS  PubMed  Google Scholar 

  6. G. U. Nienhaus, The green fluorescent protein: a key tool to study chemical processes in living cells Angew. Chem., Int. Ed. 2008 47 8992.

    Article  CAS  Google Scholar 

  7. M. Zimmer, GFP: from jellyfish to the Nobel prize and beyond Chem. Soc. Rev. 2009 38 2823.

    Article  CAS  PubMed  Google Scholar 

  8. M. V. Matz, A. F. Fradkov, Y. A. Labas, A. P. Savitsky, A. G. Zaraisky, M. L. Markelov and S. A. Lukyanov, Fluorescent proteins from nonbioluminescent Anthozoa species Nat. Biotechnol. 1999 17 969.

    Article  CAS  PubMed  Google Scholar 

  9. J. Wiedenmann, A. Schenk, C. Röcker, A. Girod, K. D. Spindler and G. U. Nienhaus, A far-red fluorescent protein with fast maturation and reduced oligomerization tendency from Entacmaea quadricolor (Anthozoa, Actinaria) Proc. Natl. Acad. Sci. U. S. A. 2002 99 11646.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. J. Wiedenmann, Die Anwendung eines fluoreszierenden Proteins und weiterer farbiger Proteine und der zugehörigen Gene aus der Artengruppe Anemonia sp. (sulcata) Pennant, (Cnidaria, Anthozoa, Actinaria) in Gentechnologie und Molekularbiologie, Offenlegungsschrift DE 197 18 640 A1, Deutsches Patent- und Markenamt, 1997, 1.

    Google Scholar 

  11. N. C. Shaner, P. A. Steinbach and R. Y. Tsien, A guide to choosing fluorescent proteins Nat. Methods 2005 2 905.

    Article  CAS  PubMed  Google Scholar 

  12. D. M. Chudakov, M. V. Matz, S. Lukyanov and K. A. Lukyanov, Fluorescent proteins and their applications in imaging living cells and tissues Physiol. Rev. 2010 90 1103.

    Article  CAS  PubMed  Google Scholar 

  13. V. Adam, B. Moeyaert, C. C. David, H. Mizuno, M. Lelimousin, P. Dedecker, R. Ando, A. Miyawaki, J. Michiels, Y. Engelborghs and J. Hofkens, Rational design of photoconvertible and biphotochromic fluorescent proteins for advanced microscopy applications Chem. Biol. 2011 18 1241.

    Article  CAS  PubMed  Google Scholar 

  14. J. Wiedenmann, S. Gayda, V. Adam, F. Oswald, K. Nienhaus, D. Bourgeois and G. U. Nienhaus, From EosFP to mIrisFP: structure-based development of advanced photoactivatable marker proteins of the GFP-family J. Biophotonics 2011 4 377.

    Article  CAS  PubMed  Google Scholar 

  15. K. A. Lukyanov, D. M. Chudakov, S. Lukyanov and V. V. Verkhusha, Innovation: Photoactivatable fluorescent proteins Nat. Rev. Mol. Cell Biol. 2005 6 885.

    Article  CAS  PubMed  Google Scholar 

  16. M. Fernandez-Suarez and A. Y. Ting, Fluorescent probes for super-resolution imaging in living cells Nat. Rev. Mol. Cell Biol. 2008 9 929.

    Article  CAS  PubMed  Google Scholar 

  17. J. Lippincott-Schwartz and G. H. Patterson, Photoactivatable fluorescent proteins for diffraction-limited and super-resolution imaging Trends Cell Biol. 2009 19 555.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. B. Huang, M. Bates and X. Zhuang, Super-resolution fluorescence microscopy Annu. Rev. Biochem. 2009 78 993.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. P. N. Hedde and G. U. Nienhaus, Optical Imaging of Nanoscale Cellular Structures Biophys. Rev. 2010 2 147.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. K. Nienhaus and G. U. Nienhaus, Fluorescent proteins for live-cell imaging with super-resolution Chem. Soc. Rev. 2014 43 1088.

    Article  CAS  PubMed  Google Scholar 

  21. S. W. Hell, Microscopy and its focal switch Nat. Methods 2009 6 24.

    Article  CAS  PubMed  Google Scholar 

  22. T. Ha and P. Tinnefeld, Photophysics of fluorescent probes for single-molecule biophysics and super-resolution imaging Annu. Rev. Phys. Chem. 2012 63 595.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. M. Zimmer, Green fluorescent protein (GFP): applications, structure, and related photophysical behavior Chem. Rev. 2002 102 759.

    Article  CAS  PubMed  Google Scholar 

  24. R. N. Day and M. W. Davidson, The fluorescent protein palette: tools for cellular imaging Chem. Soc. Rev. 2009 38 2887.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. J. Wiedenmann, F. Oswald and G. U. Nienhaus, Fluorescent proteins for live cell imaging: opportunities, limitations, and challenges IUBMB Life 2009 61 1029.

    Article  CAS  PubMed  Google Scholar 

  26. O. V. Stepanenko, D. M. Shcherbakova, I. M. Kuznetsova, K. K. Turoverov and V. V. Verkhusha, Modern fluorescent proteins: from chromophore formation to novel intracellular applications BioTechniques 2011 51 313.

    Article  PubMed  PubMed Central  Google Scholar 

  27. S. J. Remington, Fluorescent proteins: maturation, photochemistry and photophysics Curr. Opin. Struct. Biol. 2006 16 714.

    Article  CAS  PubMed  Google Scholar 

  28. C. D’Angelo, A. Denzel, A. Vogt, M. V. Matz, F. Oswald, A. Salih, G. U. Nienhaus and J. Wiedenmann, Blue light regulation of host pigment in reef-building corals Mar. Ecol.: Prog. Ser. 2008 364 97.

    Article  CAS  Google Scholar 

  29. A. G. Evdokimov, M. E. Pokross, N. S. Egorov, A. G. Zaraisky, I. V. Yampolsky, E. M. Merzlyak, A. N. Shkoporov, I. Sander, K. A. Lukyanov and D. M. Chudakov, Structural basis for the fast maturation of Arthropoda green fluorescent protein EMBO Rep. 2006 7 1006.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. T. Nagai, K. Ibata, E. S. Park, M. Kubota, K. Mikoshiba and A. Miyawaki, A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications Nat. Biotechnol. 2002 20 87.

    Article  CAS  PubMed  Google Scholar 

  31. J. Yu, J. Xiao, X. Ren, K. Lao and X. S. Xie, Probing gene expression in live cells, one protein molecule at a time Science 2006 311 1600.

    Article  CAS  PubMed  Google Scholar 

  32. S. Kredel, K. Nienhaus, M. Wolff, F. Oswald, S. Ivanchenko, F. Cymer, A. Jeromin, F. J. Michel, K.-D. Spindler, R. Heilker, G. U. Nienhaus and J. Wiedenmann, Optimized and Far-red Emitting Variants of Fluorescent Protein eqFP611 Chem. Biol. 2008 15 224.

    Article  CAS  PubMed  Google Scholar 

  33. S. Kredel, F. Oswald, K. Nienhaus, K. Deuschle, C. Röcker, M. Wolff, R. Heilker, G. U. Nienhaus and J. Wiedenmann, mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures PLoS One 2009 4 e4391.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  34. A. J. Lam, F. St-Pierre, Y. Gong, J. D. Marshall, P. J. Cranfill, M. A. Baird, M. R. McKeown, J. Wiedenmann, M. W. Davidson, M. J. Schnitzer, R. Y. Tsien and M. Z. Lin, Improving FRET dynamic range with bright green and red fluorescent proteins Nat. Methods 2012 9 1005.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. N. C. Shaner, M. Z. Lin, M. R. McKeown, P. A. Steinbach, K. L. Hazelwood, M. W. Davidson and R. Y. Tsien, Improving the photostability of bright monomeric orange and red fluorescent proteins Nat. Methods 2008 5 545.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. J. N. Henderson, M. F. Osborn, N. Koon, R. Gepshtein, D. Huppert and S. J. Remington, Excited state proton transfer in the red fluorescent protein mKeima J. Am. Chem. Soc. 2009 131 13212.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. K. D. Piatkevich, V. N. Malashkevich, S. C. Almo and V. V. Verkhusha, Engineering ESPT pathways based on structural analysis of LSSmKate red fluorescent proteins with large Stokes shift J. Am. Chem. Soc. 2010 132 10762.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. S. Violot, P. Carpentier, L. Blanchoin and D. Bourgeois, Reverse pH-dependence of chromophore protonation explains the large Stokes shift of the red fluorescent protein mKeima J. Am. Chem. Soc. 2009 131 10356.

    Article  CAS  PubMed  Google Scholar 

  39. R. Bizzarri, M. Serresi, S. Luin and F. Beltram, Green fluorescent protein based pH indicators for in vivo use: a review Anal. Bioanal. Chem. 2009 393 1107.

    Article  CAS  PubMed  Google Scholar 

  40. G. U. Nienhaus and J. Wiedenmann, Structure, dynamics and optical properties of fluorescent proteins: perspectives for marker development ChemPhysChem 2009 10 1369.

    Article  CAS  PubMed  Google Scholar 

  41. R. Ando, H. Hama, M. Yamamoto-Hino, H. Mizuno and A. Miyawaki, An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein Proc. Natl. Acad. Sci. U. S. A. 2002 99 12651.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. J. Wiedenmann, S. Ivanchenko, F. Oswald, F. Schmitt, C. Röcker, A. Salih, K. D. Spindler and G. U. Nienhaus, EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion Proc. Natl. Acad. Sci. U. S. A. 2004 101 15905.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. F. Oswald, F. Schmitt, A. Leutenegger, S. Ivanchenko, C. D’Angelo, A. Salih, S. Maslakova, M. Bulina, R. Schirmbeck, G. U. Nienhaus, M. V. Matz and J. Wiedenmann, Contributions of host and symbiont pigments to the coloration of reef corals FEBS J. 2007 274 1102.

    Article  CAS  PubMed  Google Scholar 

  44. H. Tsutsui, S. Karasawa, H. Shimizu, N. Nukina and A. Miyawaki, Semi-rational engineering of a coral fluorescent protein into an efficient highlighter EMBO Rep. 2005 6 233.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. H. Hoi, N. C. Shaner, M. W. Davidson, C. W. Cairo, J. Wang and R. E. Campbell, A monomeric photoconvertible fluorescent protein for imaging of dynamic protein localization J. Mol. Biol. 2010 401 776.

    Article  CAS  PubMed  Google Scholar 

  46. N. G. Gurskaya, V. V. Verkhusha, A. S. Shcheglov, D. B. Staroverov, T. V. Chepurnykh, A. F. Fradkov, S. Lukyanov and K. A. Lukyanov, Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light Nat. Biotechnol. 2006 24 461.

    Article  CAS  PubMed  Google Scholar 

  47. A. L. McEvoy, H. Hoi, M. Bates, E. Platonova, P. J. Cranfill, M. A. Baird, M. W. Davidson, H. Ewers, J. Liphardt and R. E. Campbell, mMaple: a photoconvertible fluorescent protein for use in multiple imaging modalities PLoS One 2012 7 e51314.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  48. V. Adam, M. Lelimousin, S. Boehme, G. Desfonds, K. Nienhaus, M. J. Field, J. Wiedenmann, S. McSweeney, G. U. Nienhaus and D. Bourgeois, Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations Proc. Natl. Acad. Sci. U. S. A. 2008 105 18343.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. H. Chang, M. Zhang, W. Ji, J. Chen, Y. Zhang, B. Liu, J. Lu, J. Zhang, P. Xu and T. Xu, A unique series of reversibly switchable fluorescent proteins with beneficial properties for various applications Proc. Natl. Acad. Sci. U. S. A. 2012 109 4455.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. S. A. McKinney, C. S. Murphy, K. L. Hazelwood, M. W. Davidson and L. L. Looger, A bright and photostable photoconvertible fluorescent protein Nat. Methods 2009 6 131.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. J. Wiedenmann and G. U. Nienhaus, Live-cell imaging with EosFP and other photoactivatable marker proteins of the GFP family Expert Rev. Proteomics 2006 3 361.

    Article  CAS  PubMed  Google Scholar 

  52. M. Zhang, H. Chang, Y. Zhang, J. Yu, L. Wu, W. Ji, J. Chen, B. Liu, J. Lu, Y. Liu, J. Zhang, P. Xu and T. Xu, Rational design of true monomeric and bright photoactivatable fluorescent proteins Nat. Methods 2012 9 727.

    Article  CAS  PubMed  Google Scholar 

  53. J. Fuchs, S. Böhme, F. Oswald, P. N. Hedde, M. Krause, J. Wiedenmann and G. U. Nienhaus, A Photoactivatable Marker Protein for Pulse-chase Imaging with Superresolution Nat. Methods 2010 7 627.

    Article  CAS  PubMed  Google Scholar 

  54. A. Schenk, S. Ivanchenko, C. Röcker, J. Wiedenmann and G. U. Nienhaus, Photodynamics of red fluorescent proteins studied by fluorescence correlation spectroscopy Biophys. J. 2004 86 384.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. A. Miyawaki, D. M. Shcherbakova and V. V. Verkhusha, Red fluorescent proteins: chromophore formation and cellular applications Curr. Opin. Struct. Biol. 2012 22 679.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. L. A. Gross, G. S. Baird, R. C. Hoffman, K. K. Baldridge and R. Y. Tsien, The structure of the chromophore within DsRed, a red fluorescent protein from coral Proc. Natl. Acad. Sci. U. S. A. 2000 97 11990.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. D. Yarbrough, R. M. Wachter, K. Kallio, M. V. Matz and S. J. Remington, Refined crystal structure of DsRed, a red fluorescent protein from coral, at 2.0-Å resolution Proc. Natl. Acad. Sci. U. S. A. 2001 98 462.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. V. V. Verkhusha, D. M. Chudakov, N. G. Gurskaya, S. Lukyanov and K. A. Lukyanov, Common pathway for the red chromophore formation in fluorescent proteins and chromoproteins Chem. Biol. 2004 11 845.

    Article  CAS  PubMed  Google Scholar 

  59. S. Pletnev, F. V. Subach, Z. Dauter, A. Wlodawer and V. V. Verkhusha, Understanding blue-to-red conversion in monomeric fluorescent timers and hydrolytic degradation of their chromophores J. Am. Chem. Soc. 2010 132 2243.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. F. V. Subach, O. M. Subach, I. S. Gundorov, K. S. Morozova, K. D. Piatkevich, A. M. Cuervo and V. V. Verkhusha, Monomeric fluorescent timers that change color from blue to red report on cellular trafficking Nat. Chem. Biol. 2009 5 118.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. O. M. Subach, V. N. Malashkevich, W. D. Zencheck, K. S. Morozova, K. D. Piatkevich, S. C. Almo and V. V. Verkhusha, Structural characterization of acylimine-containing blue and red chromophores in mTagBFP and TagRFP fluorescent proteins Chem. Biol. 2010 17 333.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. R. L. Strack, D. E. Strongin, L. Mets, B. S. Glick and R. J. Keenan, Chromophore formation in DsRed occurs by a branched pathway J. Am. Chem. Soc. 2010 132 8496.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. K. Nienhaus, H. Nar, R. Heilker, J. Wiedenmann and G. U. Nienhaus, Trans-cis isomerization is responsible for the red-shifted fluorescence in variants of the red fluorescent protein eqFP611 J. Am. Chem. Soc. 2008 130 12578.

    Article  CAS  PubMed  Google Scholar 

  64. S. Pletnev, F. V. Subach, Z. Dauter, A. Wlodawer and V. V. Verkhusha, A structural basis for reversible photoswitching of absorbance spectra in red fluorescent protein rsTagRFP J. Mol. Biol. 2012 417 144.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. F. V. Subach, V. N. Malashkevich, W. D. Zencheck, H. Xiao, G. S. Filonov, S. C. Almo and V. V. Verkhusha, Photoactivation mechanism of PAmCherry based on crystal structures of the protein in the dark and fluorescent states Proc. Natl. Acad. Sci. U. S. A. 2009 106 21097.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. S. Karasawa, T. Araki, T. Nagai, H. Mizuno and A. Miyawaki, Cyan-emitting and orange-emitting fluorescent proteins as a donor/acceptor pair for fluorescence resonance energy transfer Biochem. J. 2004 381 307.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. N. C. Shaner, R. E. Campbell, P. A. Steinbach, B. N. Giepmans, A. E. Palmer and R. Y. Tsien, Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein Nat. Biotechnol. 2004 22 1567.

    Article  CAS  PubMed  Google Scholar 

  68. T. Tsuboi, T. Kitaguchi, S. Karasawa, M. Fukuda and A. Miyawaki, Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine cells revealed by newly developed monomeric fluorescent timer protein Mol. Biol. Cell 2010 21 87.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. G. U. Nienhaus, K. Nienhaus, A. Hölzle, S. Ivanchenko, F. Renzi, F. Oswald, M. Wolff, F. Schmitt, C. Röcker, B. Vallone, W. Weidemann, R. Heilker, H. Nar and J. Wiedenmann, Photoconvertible Fluorescent Protein EosFP-Biophysical Properties and Cell Biology Applications Photochem. Photobiol. 2006 82 351.

    Article  CAS  PubMed  Google Scholar 

  70. R. E. Campbell, O. Tour, A. E. Palmer, P. A. Steinbach, G. S. Baird, D. A. Zacharias and R. Y. Tsien, A monomeric red fluorescent protein Proc. Natl. Acad. Sci. U. S. A. 2002 99 7877.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. J. Wiedenmann, B. Vallone, F. Renzi, K. Nienhaus, S. Ivanchenko, C. Röcker and G. U. Nienhaus, The red fluorescent protein eqFP611 and its genetically engineered dimeric variants J. Biomed. Opt. 2005 10 14003.

    Article  PubMed  CAS  Google Scholar 

  72. P. Abbyad, W. Childs, X. Shi and S. G. Boxer, Dynamic Stokes shift in green fluorescent protein variants Proc. Natl. Acad. Sci. U. S. A. 2007 104 20189.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. K. D. Piatkevich, V. N. Malashkevich, K. S. Morozova, N. A. Nemkovich, S. C. Almo and V. V. Verkhusha, Extended Stokes Shift in Fluorescent Proteins: Chromophore-Protein Interactions in a Near-Infrared TagRFP675 Variant Sci. Rep. 2013 3 1847.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  74. M. Z. Lin, M. R. McKeown, H. L. Ng, T. A. Aguilera, N. C. Shaner, R. E. Campbell, S. R. Adams, L. A. Gross, W. Ma, T. Alber and R. Y. Tsien, Autofluorescent proteins with excitation in the optical window for intravital imaging in mammals Chem. Biol. 2009 16 1169.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  75. D. Shcherbo, I. I. Shemiakina, A. V. Ryabova, K. E. Luker, B. T. Schmidt, E. A. Souslova, T. V. Gorodnicheva, L. Strukova, K. M. Shidlovskiy, O. V. Britanova, A. G. Zaraisky, K. A. Lukyanov, V. B. Loschenov, G. D. Luker and D. M. Chudakov, Near-infrared fluorescent proteins Nat. Methods 2010 7 827.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  76. R. A. Chica, M. M. Moore, B. D. Allen and S. L. Mayo, Generation of longer emission wavelength red fluorescent proteins using computationally designed libraries Proc. Natl. Acad. Sci. U. S. A. 2010 107 20257.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. K. S. Morozova, K. D. Piatkevich, T. J. Gould, J. Zhang, J. Bewersdorf and V. V. Verkhusha, Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy Biophys. J. 2010 99 L13.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. R. L. Strack, B. Hein, D. Bhattacharyya, S. W. Hell, R. J. Keenan and B. S. Glick, A Rapidly Maturing Far-Red Derivative of DsRed-Express2 for Whole-Cell Labeling Biochemistry 2009 48 8279.

    Article  CAS  PubMed  Google Scholar 

  79. D. C. Loos, S. Habuchi, C. Flors, J. Hotta, J. Wiedenmann, G. U. Nienhaus and J. Hofkens, Photoconversion in the red fluorescent protein from the sea anemone Entacmaea quadricolor: is cis-trans isomerization involved? J. Am. Chem. Soc. 2006 128 6270.

    Article  CAS  PubMed  Google Scholar 

  80. N. V. Pletneva, V. Z. Pletnev, I. I. Shemiakina, D. M. Chudakov, I. Artemyev, A. Wlodawer, Z. Dauter and S. Pletnev, Crystallographic study of red fluorescent protein eqFP578 and its far-red variant Katushka reveals opposite pH-induced isomerization of chromophore Protein Sci. 2011 20 1265.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. X. Shu, N. C. Shaner, C. A. Yarbrough, R. Y. Tsien and S. J. Remington, Novel chromophores and buried charges control color in mFruits Biochemistry 2006 45 9639.

    Article  CAS  PubMed  Google Scholar 

  82. M. Drobizhev, S. Tillo, N. S. Makarov, T. E. Hughes and A. Rebane, Color hues in red fluorescent proteins are due to internal quadratic Stark effect J. Phys. Chem. B 2009 113 12860.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. R. M. Wachter, M. A. Elsliger, K. Kallio, G. T. Hanson and S. J. Remington, Structural basis of spectral shifts in the yellow-emission variants of green fluorescent protein Structure 1998 6 1267.

    Article  CAS  PubMed  Google Scholar 

  84. J. Akerboom, N. Carreras Calderon, L. Tian, S. Wabnig, M. Prigge, J. Tolo, A. Gordus, M. B. Orger, K. E. Severi, J. J. Macklin, R. Patel, S. R. Pulver, T. J. Wardill, E. Fischer, C. Schuler, T. W. Chen, K. S. Sarkisyan, J. S. Marvin, C. I. Bargmann, D. S. Kim, S. Kugler, L. Lagnado, P. Hegemann, A. Gottschalk, E. R. Schreiter and L. L. Looger, Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics Front. Mol. Neurosci. 2013 6 2.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. F. V. Subach, K. D. Piatkevich and V. V. Verkhusha, Directed molecular evolution to design advanced red fluorescent proteins Nat. Methods 2011 8 1019.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. P. Konold, C. K. Regmi, P. P. Chapagain, B. S. Gerstman and R. Jimenez, Hydrogen bond flexibility correlates with Stokes shift in mPlum variants J. Phys. Chem. B 2014 118 2940.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  87. D. Shcherbo, E. M. Merzlyak, T. V. Chepurnykh, A. F. Fradkov, G. V. Ermakova, E. A. Solovieva, K. A. Lukyanov, E. A. Bogdanova, A. G. Zaraisky, S. Lukyanov and D. M. Chudakov, Bright far-red fluorescent protein for whole-body imaging Nat. Methods 2007 4 741.

    Article  CAS  PubMed  Google Scholar 

  88. B. J. Bevis and B. S. Glick, Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed) Nat. Biotechnol. 2002 20 83.

    Article  CAS  PubMed  Google Scholar 

  89. S. A. Lukyanov, and D. M. Chudakov, Novel fluorescent proteins and methods for using same, Patent US 20100015701 A1, 2007.

    Google Scholar 

  90. L. Wang, W. C. Jackson, P. A. Steinbach and R. Y. Tsien, Evolution of new nonantibody proteins via iterative somatic hypermutation Proc. Natl. Acad. Sci. U. S. A. 2004 101 16745.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Applied Physics and Center for Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), Wolfgang-Gaede-Strasse 1, 76131, Karlsruhe, Germany

    Anika Hense, Karin Nienhaus & G. Ulrich Nienhaus

  2. Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, 76344, Eggenstein-Leopoldshafen, Germany

    G. Ulrich Nienhaus

  3. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois, 61801, USA

    G. Ulrich Nienhaus

Authors
  1. Anika Hense
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Karin Nienhaus
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Ulrich Nienhaus
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. Ulrich Nienhaus.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hense, A., Nienhaus, K. & Nienhaus, G.U. Exploring color tuning strategies in red fluorescent proteins. Photochem Photobiol Sci 14, 200–212 (2015). https://doi.org/10.1039/c4pp00212a

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1039/c4pp00212a

Search

Navigation