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Identification of GFP-like Proteins in Nonbioluminescent, Azooxanthellate Anthozoa Opens New Perspectives for Bioprospecting

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Abstract

We screened nonbioluminescent, azooxanthellate cnidaria as potential sources for advanced marker proteins and succeeded in cloning a tetrameric green fluorescent protein (GFP) from the tentacles of Cerianthus membranaceus. The fluorescence of this protein (cmFP512) is characterized by excitation maximum at 503 nm, emission maximum at 512 nm, extinction coefficient of 58,800 M−1 cm−1, quantum yield of 0.66, and fluorescence lifetime of 2.4 ns. The chromophore is formed from the tripeptide Gln-Tyr-Gly. The amino acid sequence of this protein shares 17.8% identical residues with GFP from Aequorea victoria. Weak interactions between the subunits of the tetramer make cmFP512 a promising lead structure for the generation of monomeric variants of fluorescent proteins. Both red fluorescent proteins and nonfluorescent proteins of the GFP family were also purified from tissue homogenates of Adamsia palliata and Calliactis parasitica. The results presented here indicate that a photoprotective function of GFP-like proteins is unlikely in the examined anthozoa species.

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References

  • R. Ando H. Hama M. Yamamoto-Hino H. Mizuno A. Miyawaki (2002) ArticleTitleAn optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein Proc Natl Acad Sci U S A 99 12651–12656 Occurrence Handle10.1073/pnas.202320599 Occurrence Handle1:CAS:528:DC%2BD38XnvFGhsrs%3D Occurrence Handle12271129

    Article  CAS  PubMed  Google Scholar 

  • G.S. Baird D.A. Zacharias R.Y. Tsien (2000) ArticleTitleBiochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral Proc Natl Acad Sci U S A 97 11984–11989 Occurrence Handle10.1073/pnas.97.22.11984 Occurrence Handle1:CAS:528:DC%2BD3cXnvVSgtro%3D Occurrence Handle11050229

    Article  CAS  PubMed  Google Scholar 

  • B. Bevis B. Glick (2002) ArticleTitleRapidly maturing variants of Discosoma red fluorescent protein (DsRED) Nat Biotechnol 20 83–86 Occurrence Handle10.1038/nbt0102-83 Occurrence Handle1:CAS:528:DC%2BD38XislSkuw%3D%3D Occurrence Handle11753367

    Article  CAS  PubMed  Google Scholar 

  • N. Billinton A.W. Knight (2001) ArticleTitleSeeing the wood through the trees: a review of techniques for distinguishing green fluorescent protein from endogenous autofluorescence Anal Biochem 291 175–197 Occurrence Handle10.1006/abio.2000.5006 Occurrence Handle1:CAS:528:DC%2BD3MXitlaqu7s%3D Occurrence Handle11401292

    Article  CAS  PubMed  Google Scholar 

  • R.E. Campbell O. Tour A.E. Palmer PA. Steinbach G.S. Baird D.A. Zacharias R.Y. Tsien (2002) ArticleTitleA monomeric red fluorescent protein Proc Natl Acad Sci U S A 99 7877–7882 Occurrence Handle10.1073/pnas.082243699 Occurrence Handle1:CAS:528:DC%2BD38XkvVGjsbw%3D Occurrence Handle12060735

    Article  CAS  PubMed  Google Scholar 

  • M. Chalfie Y. Tu G. Euskirchen W.W. Ward D.C. Prasher (1994) ArticleTitleGreen fluorescent proteins as a marker for gene expression Science 263 802–805 Occurrence Handle1:CAS:528:DyaK2cXitFWkurc%3D Occurrence Handle8303295

    CAS  PubMed  Google Scholar 

  • S.K. Deo S. Daunert (2001) ArticleTitleSpecial issue paper: luminescent proteins from Aequorea victoria: applications in drug discovery and in high throughput analysis Fresenius J Anal Chem 369 258–266 Occurrence Handle10.1007/s002160000646 Occurrence Handle1:CAS:528:DC%2BD3MXhtFamu7c%3D Occurrence Handle11293702

    Article  CAS  PubMed  Google Scholar 

  • S.G. Dove O. Hoegh-Guldberg S. Ranganathan (2001) ArticleTitleMajor colour patterns of reef-building are due to a family of GFP-like proteins Coral Reefs 19 197–204

    Google Scholar 

  • M. Edmunds G.W. Potts R.C. Swinfen V.L. Waters (1976) ArticleTitleDefensive behaviour of sea anemones in response to predation by the ophistobranch mollusc Aeolidia papillosa (L) J Mar Biol Assoc UK 54 939–947

    Google Scholar 

  • A.F. Fradkov Y. Chen L. Ding E.V. Barsova M.V. Matz S.A. Lukyanov (2000) ArticleTitleNovel fluorescent protein from Discosoma coral and its mutants possesses a unique far-red fluorescence FEBS Lett 479 127–130 Occurrence Handle10.1016/S0014-5793(00)01895-0 Occurrence Handle1:CAS:528:DC%2BD3cXls1Gisbs%3D Occurrence Handle10981720

    Article  CAS  PubMed  Google Scholar 

  • D.L. Fox C.F.A. Pantin (1941) ArticleTitleThe colours of the plumose anemone Metridium senile (L.) Phil Trans R Soc Lond 230 415–450

    Google Scholar 

  • D.L. Fox D.W. Wilkie F.T. Haxo (1978) ArticleTitleCarotenoid fractionation in the plumose anemone Metridium II: search for dietary sources of ovarian astaxanthin Comp Biochem Physiol 59B 289–294 Occurrence Handle1:CAS:528:DyaE1cXlsVemtLY%3D

    CAS  Google Scholar 

  • S.C. Gill P.H. von Hippel (1989) ArticleTitleCalculation of protein extinction coefficients from amino acid sequence data Anal Biochem 182 319–326 Occurrence Handle1:CAS:528:DyaL1MXmt1ygt7Y%3D Occurrence Handle2610349

    CAS  PubMed  Google Scholar 

  • N.G. Gurskaya A.F. Fradkov A. Terskikh MV. Matz Y.A. Labas V.I. Martynov Y.G. Yanushevich K.A. Lukyanov S.A. Lukyanov (2001) ArticleTitleGFP-like chromoproteins as a source of far-red fluorescent proteins FEBS Lett 507 16–20 Occurrence Handle10.1016/S0014-5793(01)02930-1 Occurrence Handle1:CAS:528:DC%2BD3MXnslWrtrs%3D Occurrence Handle11682051

    Article  CAS  PubMed  Google Scholar 

  • V. Häusermann (2003) Ordnung Ceriantharia (Zylinderrosen) R. Hofrichter (Eds) Das Mittelmeer, Fauna, Flora, Ökologie Spektrum Akademischer Verlag Heidelberg, Germany 450–453

    Google Scholar 

  • S. Hertzberg S. Liaaen-Jensen C.R. Enzell G.W. Francis (1969) ArticleTitleAnimal carotinoids 3: the carotenoids of Actinia equina—structure determination of actinioerythrin and violerythrin Acta Chem Scand 23 3290–3312 Occurrence Handle1:CAS:528:DyaE3cXhtVSqsb4%3D

    CAS  Google Scholar 

  • S. Kawaguti (1969) ArticleTitleEffect of the green fluorescent pigment on the productivity of reef corals Micronesia 5 IssueID2 313

    Google Scholar 

  • Y.A. Labas N.G. Gurskaya Y.G. Yanushevich A.F. Fradkov K.A. Lukyanov S.A. Lukyanov M.V. Matz (2002) ArticleTitleDiversity and evolution of the green fluorescent protein family Proc Natl Acad Sci U S A 99 4256–4261 Occurrence Handle10.1073/pnas.062552299 Occurrence Handle1:CAS:528:DC%2BD38XivFShsbg%3D Occurrence Handle11929996

    Article  CAS  PubMed  Google Scholar 

  • R.D. LeBoeuf S.A. McCommas N.R. Howe J.D. Tauber (1981) ArticleTitleThe role of carotenoids in the colour polymorphism of the sea anemone, Bunodosoma granulifera (Anthozoa: Actinaria) Comp Biochem Physiol 68B 25–29 Occurrence Handle1:CAS:528:DyaL3MXhslWhtL0%3D

    CAS  Google Scholar 

  • K.A. Lukyanov A.F. Fradkov N.G. Gurskaya M.V. Matz Y.A. Labas A.P. Savitsky M.L. Markelov A.G. Zaraisky X. Zhao Y. Fang W. Tan S.A. Lukyanov (2000) ArticleTitleNatural animal coloration can be determind by a non-fluorescent green fluorescent protein homolog J Biol Chem 275 25879–25882 Occurrence Handle10.1074/jbc.C000338200 Occurrence Handle1:CAS:528:DC%2BD3cXmt1OlurY%3D Occurrence Handle10852900

    Article  CAS  PubMed  Google Scholar 

  • V.I. Martynov A.P. Savitsky N.Y. Martynova P.A. Savitsky K.A. Lukyanov S.A. Lukyanov (2001) ArticleTitleAlternative cyclization in GFP-like proteins family: the formation and structure of the chromophore of a purple chromoprotein from Anemonia sulcata J Biol Chem 276 21012–21016 Occurrence Handle10.1074/jbc.M100500200 Occurrence Handle1:CAS:528:DC%2BD3MXksFGlsLY%3D Occurrence Handle11259412

    Article  CAS  PubMed  Google Scholar 

  • M.V. Matz A.F. Fradkov Y.A. Labas A.P. Savitsky A.G. Zaraisky M.L. Markelov S.A. Lukyanov (1999) ArticleTitleFluorescent proteins from non-bioluminescent Anthozoa species Nat Biotechnol 17 969–973 Occurrence Handle10.1038/13657 Occurrence Handle1:CAS:528:DyaK1MXmvVelsrk%3D Occurrence Handle10504696

    Article  CAS  PubMed  Google Scholar 

  • K. Nienhaus B. Vallone F. Renzi J. Wiedenmann G.U. Nienhaus (2003) ArticleTitleCrystallization and preliminary X-ray diffraction analysis of the red fluorescent protein FP611 Acta Cryst D59 1253–1255 Occurrence Handle1:CAS:528:DC%2BD3sXkvFSktrg%3D

    CAS  Google Scholar 

  • J. Olmsted (1979) ArticleTitleCalorimetric determinations of absolute fluorescence quantum yields J Phys Chem 83 2581–2584 Occurrence Handle1:CAS:528:DyaE1MXls1Gkurs%3D

    CAS  Google Scholar 

  • F. Pax I. Müller (1962) ArticleTitleDie Anthozoenfauna der Adria Split: Institut für Ozeanographie und Fischerei . .

    Google Scholar 

  • R. Pepperkok A. Squire S. Geley P.I. Bastiaens (1999) ArticleTitleSimultaneous detection of multiple green fluorescent proteins in live cells by fluorescence lifetime imaging microscopy Curr Biol 9 269–272 Occurrence Handle10.1016/S0960-9822(99)80117-1 Occurrence Handle1:CAS:528:DyaK1MXhvFejtrc%3D Occurrence Handle10074454

    Article  CAS  PubMed  Google Scholar 

  • D.C. Prasher V.K. Eckenrode W.W. Ward F.G. Prendergast MJ. Cormier (1992) ArticleTitlePrimary structure of the Aeqourea victoria green fluorescent protein Gene 111 229–233 Occurrence Handle10.1016/0378-1119(92)90691-H Occurrence Handle1:CAS:528:DyaK3sXitVOktLw%3D Occurrence Handle1347277

    Article  CAS  PubMed  Google Scholar 

  • A. Salih A. Larkum G. Cox M. Kühl O. Hoegh-Guldberg (2000) ArticleTitleFluorescent pigments in corals are photoprotective Nature 408 850–853 Occurrence Handle10.1038/35048564 Occurrence Handle1:CAS:528:DC%2BD3MXhtlaisA%3D%3D Occurrence Handle11130722

    Article  CAS  PubMed  Google Scholar 

  • C.S. Szent-Gyorgyi B.J. Bryan (2001) ArticleTitleLuciferases, fluorescent proteins, nucleic acids encoding the luciferases and fluorescent proteins and the use thereof in diagnostics, high throughput screening and novelty items US Patent 6232107-B . .

    Google Scholar 

  • RY Tsien (1998) ArticleTitleThe green fluorescent protein Annu Rev Biochem 67 509–544 Occurrence Handle10.1146/annurev.biochem.67.1.509 Occurrence Handle1:CAS:528:DyaK1cXlsFOmsb8%3D Occurrence Handle9759496

    Article  CAS  PubMed  Google Scholar 

  • W.W. Ward (1998) Biochemical and physical properties of green fluorescent protein M. Chalfie S. Kain (Eds) Green Fuorescent Protein: Properties, Applications, and Protocols Wiley-Liss New York, N.Y. 45–75

    Google Scholar 

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

    Google Scholar 

  • J. Wiedemann C. Röcker W. Funke (1999) The morphs of Anemonia aff. sulcata (Cnidaria, Anthozoa) in particular consideration of the ectodermal pigments J. Pfadenhauer (Eds) Verhandlungen der Gesellschaft für Ökologie, Band 29 Spektrum Akademischer Verlag Heidelberg, Germany 497–503

    Google Scholar 

  • J. Wiedenmann C. Elke K-D. Spindler W. Funke (2000) ArticleTitleCracks in the β-can: fluorescent proteins from Anemonia sulcata Proc Natl Acad Sci U S A 97 14091–14096 Occurrence Handle10.1073/pnas.97.26.14091 Occurrence Handle1:CAS:528:DC%2BD3MXitVChtw%3D%3D Occurrence Handle11121018

    Article  CAS  PubMed  Google Scholar 

  • J. Wiedemann A. Schenk C. Röcker K.-D. Spindler G.U. Nienhaus (2002) ArticleTitleA far-red fluorescent protein with fast maturation and reduced oligomerization tendency from Entacmaea quadricolor (Cnidaria, Anthozoa, Actinaria) Proc Natl Acad Sci U S A 99 11646–11651 Occurrence Handle10.1073/pnas.182157199 Occurrence Handle12185250

    Article  PubMed  Google Scholar 

  • R. William M.P. Ware R.K. Bauer (1983) ArticleTitlePerfomance characteristics of a small side-window photomultiplier in laser single-photon fluorescence decay measurements Rev Sci Instruments 54 1148–1156 Occurrence Handle10.1063/1.1137540

    Article  Google Scholar 

  • Y.G. Yanushevich D.B. Staroverov A.P. Savitsky A.F. Fradkov N.G. Gurskaya M.E. Bulina K.A. Lukyanov S.A. Lukyanov (2002) ArticleTitleA strategy for the generation of non-aggregating mutants of Anthozoa fluorescent proteins FEBS Lett 511 11–14 Occurrence Handle10.1016/S0014-5793(01)03263-X Occurrence Handle1:CAS:528:DC%2BD38XovFenug%3D%3D Occurrence Handle11821040

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We acknowledge the skillfull technical assistance of Ms. Maja Lenz and Mr. Uwe Theilen. We thank Ms. Marie Mühl for dedicated organizational support and Dr. Charly Röcker for helpful discussions. The work was financed in part by the University of Ulm (Anfangsförderung to J.W.), the Landesstiftung Baden-Württemberg (Elite-Postdoc-Förderung to J.W.), and the Deutsche Forschungsgemeinschaft (SFB 569 and GRK 328 to G.U.N.).

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Authors and Affiliations

  1. Department of General Zoology and Endocrinology, University of Ulm, 89069, Ulm, Germany

    Jörg Wiedenmann

  2. Department of Biophysics, University of Ulm, 89069, Ulm, Germany

    Sergey Ivanchenko & G. Ulrich. Nienhaus

  3. Department of Internal Medicine I, University of Ulm, 89069, Ulm, Germany

    Franz Oswald

  4. Department of Physics, University of Illinois at Urbana–Champaign, Urbana, IL, 61801, U.S.A

    G. Ulrich. Nienhaus

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  1. Jörg Wiedenmann
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  2. Sergey Ivanchenko
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  3. Franz Oswald
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  4. G. Ulrich. Nienhaus
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Correspondence to Jörg Wiedenmann.

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Wiedenmann, J., Ivanchenko, S., Oswald, F. et al. Identification of GFP-like Proteins in Nonbioluminescent, Azooxanthellate Anthozoa Opens New Perspectives for Bioprospecting. Mar. Biotechnol. 6, 270–277 (2004). https://doi.org/10.1007/s10126-004-3006-4

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