Skip to main content

Advertisement

SpringerLink
Log in

Large-scale Expression and Purification of a G-protein-coupled Receptor for Structure Determination – An Overview

  • Published:
Journal of Structural and Functional Genomics

Abstract

Structure determination of G-protein-coupled receptors and other applications, such as nuclear magnetic resonance studies, require milligram quantities of purified, functional receptor protein on a regular basis. We present an overview on expression and purification studies with a receptor for neurotensin. Functional expression in Escherichia coli and an automated two-column purification routine allow ongoing crystallization experiments and studies on receptor-bound ligands.

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

Abbreviations

GPCR:

G-protein-coupled receptor

H10:

deca-histidine tag

MBP:

E. coli maltose-bind- ing protein

NTR:

rat neurotensin receptor NTS1

TrxA:

E. coli thioredoxin

References

  1. T.H. Ji M. Grossmann I. Ji (1998) J. Biol. Chem. 273 17299–17302 Occurrence Handle10.1074/jbc.273.28.17299 Occurrence Handle9651309

    Article  PubMed  Google Scholar 

  2. U. Gether B.K. Kobilka (1998) J. Biol. Chem. 273 17979–17982 Occurrence Handle10.1074/jbc.273.29.17979 Occurrence Handle9660746

    Article  PubMed  Google Scholar 

  3. D.K. Vassilatis et al. (2003) Proc. Natl. Acad. Sci. USA 100 4903–4908 Occurrence Handle10.1073/pnas.0230374100 Occurrence Handle12679517

    Article  PubMed  Google Scholar 

  4. J.M. Baldwin G.F.X. Schertler V.M. Unger (1997) J. Mol. Biol. 272 144–164 Occurrence Handle10.1006/jmbi.1997.1240 Occurrence Handle9299344

    Article  PubMed  Google Scholar 

  5. T. Okada Y. Fujiyoshi M. Silow J. Navarro E.M. Landau Y. Shichida (2002) Proc. Natl. Acad. Sci. USA 99 5982–5987 Occurrence Handle10.1073/pnas.082666399 Occurrence Handle11972040

    Article  PubMed  Google Scholar 

  6. K. Palczewski et al. (2000) Science 289 739–745 Occurrence Handle10.1126/science.289.5480.739 Occurrence Handle10926528

    Article  PubMed  Google Scholar 

  7. H. Inooka et al. (2001) Nat. Struct. Biol. 8 161–165 Occurrence Handle10.1038/84159 Occurrence Handle11175907

    Article  PubMed  Google Scholar 

  8. S. Luca J.F. White A.K. Sohal D.V. Filippov J.H. Boom Particlevan R. Grisshammer M. Baldus (2003) Proc. Natl. Acad. Sci. USA 100 10706–10711 Occurrence Handle10.1073/pnas.1834523100 Occurrence Handle12960362

    Article  PubMed  Google Scholar 

  9. P.T.F. Williamson S. Bains C. Chung R. Cooke A. Watts (2002) Febs Lett. 518 111–115 Occurrence Handle10.1016/S0014-5793(02)02656-X Occurrence Handle11997028

    Article  PubMed  Google Scholar 

  10. P.J. Reeves J. Klein-Seetharaman E.V. Getmanova M. Eilers M.C. Loewen S.O. Smith H.G. Khorana (1999) Biochem. Soc. Trans. 27 950–955 Occurrence Handle10830134

    PubMed  Google Scholar 

  11. H. Furukawa et al. (2002) Mol. Pharmacol. 62 778–787 Occurrence Handle10.1124/mol.62.4.778 Occurrence Handle12237324

    Article  PubMed  Google Scholar 

  12. J.-P. Vincent J. Mazella P. Kitabgi (1999) Trends Pharmacol. Sci. 20 302–309 Occurrence Handle10.1016/S0165-6147(99)01357-7 Occurrence Handle10390649

    Article  PubMed  Google Scholar 

  13. R. Carraway S.E. Leeman (1973) J. Biol. Chem. 248 6854–6861 Occurrence Handle4745447

    PubMed  Google Scholar 

  14. K. Tanaka M. Masu S. Nakanishi (1990) Neuron 4 847–854 Occurrence Handle10.1016/0896-6273(90)90137-5 Occurrence Handle1694443

    Article  PubMed  Google Scholar 

  15. Issue on Expression and Purification of Membrane Proteins (1999) Biochem. Soc. Trans. 27, 883–960

    Google Scholar 

  16. Special Issue on Overexpression of Integral Membrane Proteins (2003) Biochim. Biophys. Acta 1610, 1–153

    Google Scholar 

  17. R. Grisshammer C.G. Tate (1995) Quart. Rev. Biophys. 28 315–422

    Google Scholar 

  18. Reinhart, C., Weiss, H.M. and Reiländer, H. (2002) In A Practical Guide to Membrane Protein Purification (Eds., Hunte, C., von Jagow, G. and Schägger, H.), Academic Press, pp. 167–178

  19. H. Reiländer C. Reinhart A. Szmolenszky (1999) NoChapterTitle T. Haga G. Berstein (Eds) G-Protein-Coupled Receptors CRC Press Boca Raton 281–322

    Google Scholar 

  20. A. Lala H.T. Sojar E. DeNardin (1997) Biochem. Pharmacol. 54 381–390 Occurrence Handle10.1016/S0006-2952(97)00191-3 Occurrence Handle9278097

    Article  PubMed  Google Scholar 

  21. J.-L. Banères A. Martin P. Hullot J.-P. Girard J.-C. Rossi J. Parello (2003) J. Mol. Biol. 329 801–814 Occurrence Handle10.1016/S0022-2836(03)00438-8 Occurrence Handle12787679

    Article  PubMed  Google Scholar 

  22. H. Kiefer R. Vogel K. Maier (2000) Receptors & Channels 7 109–119

    Google Scholar 

  23. H. Kiefer K. Maier R. Vogel (1999) Biochem. Soc. Trans. 27 908–912 Occurrence Handle10830126

    PubMed  Google Scholar 

  24. R. Grisshammer R. Duckworth R. Henderson (1993) Biochem. J. 295 571–576 Occurrence Handle8240259

    PubMed  Google Scholar 

  25. B. Bertin M. Freissmuth R.M. Breyer W. Schütz A.D. Strosberg S. Marullo (1992) J. Biol. Chem. 267 8200–8206 Occurrence Handle1533220

    PubMed  Google Scholar 

  26. G. Heijne Particlevon (1989) Nature 341 456–458 Occurrence Handle10.1038/341456a0 Occurrence Handle2677744

    Article  PubMed  Google Scholar 

  27. D. Drew L. Fröderberg L. Baars J.-W.L. Gier Particlede (2003) Biochim. Biophys. Acta 1610 3–10 Occurrence Handle12586374

    PubMed  Google Scholar 

  28. J. Tucker R. Grisshammer (1996) Biochem. J. 317 891–899 Occurrence Handle8760379

    PubMed  Google Scholar 

  29. R. Grisshammer E. Hermans (2001) Febs Lett. 493 101–105 Occurrence Handle10.1016/S0014-5793(01)02281-5 Occurrence Handle11287004

    Article  PubMed  Google Scholar 

  30. J.F. White L.B. Trinh J. Shiloach R. Grisshammer (2004) Febs Lett. 564 289–293 Occurrence Handle10.1016/S0014-5793(04)00195-4 Occurrence Handle15111111

    Article  PubMed  Google Scholar 

  31. B. Calandra J. Tucker D. Shire R. Grisshammer (1997) Biotechnol. Lett. 19 425–428 Occurrence Handle10.1023/A:1018383808401

    Article  Google Scholar 

  32. H.M. Weiß R. Grisshammer (2002) Eur. J. Biochem. 269 82–92 Occurrence Handle10.1046/j.0014-2956.2002.02618.x Occurrence Handle11784301

    Article  PubMed  Google Scholar 

  33. E.C. Hulme C.A.M. Curtis (1998) Biochem. Soc. Trans. 26 S361 Occurrence Handle10047875

    PubMed  Google Scholar 

  34. L. Stanasila D. Massotte B.L. Kieffer F. Pattus (1999) Eur. J. Biochem. 260 430–438 Occurrence Handle10.1046/j.1432-1327.1999.00187.x Occurrence Handle10095778

    Article  PubMed  Google Scholar 

  35. G. Münch P. Walker J. Shine H. Herzog (1995) Receptors Channels 3 291–297 Occurrence Handle8834002

    PubMed  Google Scholar 

  36. W. Hampe R.-H. Voss W. Haase F. Boege H. Michel H. Reiländer (2000) J. Biotechnol. 77 219–234 Occurrence Handle10.1016/S0168-1656(99)00216-3 Occurrence Handle10682281

    Article  PubMed  Google Scholar 

  37. R. Grisshammer J. Little D. Aharony (1994) Receptors Channels 2 295–302 Occurrence Handle7719707

    PubMed  Google Scholar 

  38. H. Furukawa T. Haga (2000) J. Biochem. (Tokyo) 127 151–161

    Google Scholar 

  39. S.H. White (2003) Febs Lett. 555 116–121 Occurrence Handle10.1016/S0014-5793(03)01153-0 Occurrence Handle14630330

    Article  PubMed  Google Scholar 

  40. T. Warne J. Chirnside G.F.X. Schertler (2003) Biochim. Biophys. Acta 1610 133–140 Occurrence Handle12586387

    PubMed  Google Scholar 

  41. P.D. Laible H.N. Scott L. Henry D.K. Hanson (2004) J. Struct. Funct. Genomics 5 167–172 Occurrence Handle10.1023/B:JSFG.0000029201.33710.46 Occurrence Handle15263855

    Article  PubMed  Google Scholar 

  42. P.J. Reeves J.-M. Kim H.G. Khorana (2002) Proc. Natl. Acad. Sci. 99 13413–13418 Occurrence Handle10.1073/pnas.212519199 Occurrence Handle12370422

    Article  PubMed  Google Scholar 

  43. T. Ohtaki et al. (1998) J. Biol. Chem. 273 15464–15473 Occurrence Handle10.1074/jbc.273.25.15464 Occurrence Handle9624132

    Article  PubMed  Google Scholar 

  44. C.H.W. Klaassen P.H.M. Bovee-Geurts G.L.J. DeCaluwé W.J. Grip ParticleDe (1999) Biochem. J. 342 293–300 Occurrence Handle10.1042/0264-6021:3420293 Occurrence Handle10455014

    Article  PubMed  Google Scholar 

  45. V.R.P. Ratnala H.G.P. Swarts J. Oostrum ParticleVan R. Leurs H.J.M. Groot ParticleDe R.A. Bakker W.J. Grip ParticleDe (2004) Eur. J. Biochem. 271 2636–2646 Occurrence Handle10.1111/j.1432-1033.2004.04192.x Occurrence Handle15206929

    Article  PubMed  Google Scholar 

  46. R. Grisshammer J. Tucker (1997) Protein Expres. Purif. 11 53–60 Occurrence Handle10.1006/prep.1997.0766

    Article  Google Scholar 

  47. R. Grisshammer P. Averbeck A.K. Sohal (1999) Biochem. Soc. Trans. 27 899–903 Occurrence Handle10830124

    PubMed  Google Scholar 

  48. A.K. Mohanty C.R. Simmons M.C. Wiener (2003) Protein Expres. Purif. 27 109–114 Occurrence Handle10.1016/S1046-5928(02)00589-2

    Article  Google Scholar 

  49. D.A. Daniels A.K. Sohal S. Rees R. Grisshammer (2002) Anal. Biochem. 305 214–226 Occurrence Handle10.1006/abio.2002.5663 Occurrence Handle12054450

    Article  PubMed  Google Scholar 

  50. Reprinted by permission of Federation of the European Biochemical Societies from ‘Automated large-scale purification of a G-protein-coupled receptor for neurotensin’, by White et al., Febs Lett. 564, 289–293, 2004

Download references

Author information

Authors and Affiliations

  1. Laboratory of Molecular Biology of the National Institute of Diabetes and Digestive and Kidney Diseases, Department of Health and Human Services, National Institutes of Health, Bethesda, Maryland, 20892, USA

    Reinhard Grisshammer & Jim F. White

  2. Biotechnology Unit of the National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Department of Health and Human Services, National Institutes of Health, Bethesda, Maryland, 20892, USA

    Loc B. Trinh & Joseph Shiloach

Authors
  1. Reinhard Grisshammer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jim F. White
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Loc B. Trinh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Joseph Shiloach
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Reinhard Grisshammer.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Grisshammer, R., White, J.F., Trinh, L.B. et al. Large-scale Expression and Purification of a G-protein-coupled Receptor for Structure Determination – An Overview. J Struct Funct Genomics 6, 159–163 (2005). https://doi.org/10.1007/s10969-005-1917-6

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10969-005-1917-6

Key words

Search

Navigation