Skip to main content
SpringerLink
Log in

Crystallization of Membrane Proteins

  • Protocol
  • First Online:
Membrane Biogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1033))

Abstract

The crystallization of membrane proteins is an essential technique for the determination of atomic models of three-dimensional structures by X-ray crystallography. The compositions of solutions of purified membrane proteins are altered, so as to transiently induce supersaturation, a requirement for crystal nucleation and growth. The establishment of the precise optimal crystallization conditions has to be performed individually by a combination of systematic approaches and trial-and-error. These procedures have become more efficient due to the introduction of laboratory automation. Here we describe the crystallization of the dihaem-containing quinol:fumarate reductase (QFR) membrane protein complex and illustrate key factors important in the screening process.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Deisenhofer J, Epp O, Miki K, Huber R, Michel H (1985) Structure of the protein subunits in the photosynthetic reaction center of Rhodopseudomonas viridis at 3Å resolution. Nature 318:618–624

    Article  PubMed  CAS  Google Scholar 

  2. Kendrew JC, Bodo G, Dintzis HM, Parrish RG, Wyckoff H, Phillips DC (1958) 3-Dimensional model of the myoglobin molecule obtained by X-ray analysis. Nature 181:662–666

    Article  PubMed  CAS  Google Scholar 

  3. Bergfors TM (2009) Protein crystallization, 2nd edn. International University Line, La Jolla

    Google Scholar 

  4. McPherson A (1990) Current approaches to macromolecular crystallization. Eur J Biochem 189:1–23

    Article  PubMed  CAS  Google Scholar 

  5. Watson AA, O’Callaghan CA (2005) Crystallization and X-ray diffraction analysis of human CLEC-2. Acta Crystallogr Sect F Struct Biol Cryst Commun 61:1094–1096

    Article  PubMed  CAS  Google Scholar 

  6. Michel H (ed) (1990) Crystallization of membrane proteins. CRC, Boca Raton

    Google Scholar 

  7. Hunte C, von Jagow G, Schägger H (2003) Membrane protein purification and crystallization. A practical guide, 2nd edn. Academic, San Diego

    Google Scholar 

  8. Iwata S (ed) (2003) Methods and results in the crystallization of membrane proteins. International University Line, La Jolla

    Google Scholar 

  9. Privé GG (2007) Detergents for the stabilization and crystallization of membrane proteins. Methods 41:388–397

    Article  PubMed  Google Scholar 

  10. Schulz GE (2011) A new classification of membrane protein crystals. J Mol Biol 407: 640–646

    Article  PubMed  CAS  Google Scholar 

  11. Michel H (1983) Crystallization of membrane proteins. Trends Biochem Sci 8:56–59

    Article  CAS  Google Scholar 

  12. Caffrey M (2003) Membrane protein crystallization. J Struct Biol 142:108–132

    Article  PubMed  CAS  Google Scholar 

  13. Hunte C, Michel H (2002) Crystallisation of membrane proteins mediated by antibody fragments. Curr Opin Struct Biol 12:503–508

    Article  PubMed  CAS  Google Scholar 

  14. Lieberman RL, Culver JA, Entzminger KC, Pai JC, Maynard JA (2011) Crystallization chaperone strategies for membrane proteins. Methods 55:293–302

    Article  PubMed  CAS  Google Scholar 

  15. Michel H (1982) 3-Dimensional crystals of a membrane protein complex—the photosynthetic reaction center from Rhodopseudomonas viridis. J Mol Biol 158:567–572

    Article  PubMed  CAS  Google Scholar 

  16. Gast P, Hemelrijk P, Hoff AJ (1994) Determination of the number of detergent molecules associated with the reaction center protein isolated from the photosynthetic bacterium Rhodopseudomonas viridis—effects of the amphiphilic molecule 1,2,3-heptanetriol. FEBS Lett 337:39–42

    Article  PubMed  CAS  Google Scholar 

  17. Koepke J, Hu XC, Muenke C, Schulten K, Michel H (1996) The crystal structure of the light-harvesting complex II (B800-850) from Rhodospirillum molischianum. Structure 4: 581–597

    Article  PubMed  CAS  Google Scholar 

  18. Palczewski K, Kumasaka T, Hori T, Behnke CA, Motoshima H, Fox BA, Le Trong I, Teller DC, Okada T, Stenkamp RE, Yamamoto M, Miyano M (2000) Crystal structure of rhodopsin: a G protein-coupled receptor. Science 289:739–745

    Article  PubMed  CAS  Google Scholar 

  19. Schertler GFX, Bartunik HD, Michel H, Oesterhelt D (1993) Orthorhombic crystal form of bacteriorhodopsin nucleated on benzamidine diffracting to 3.6Å resolution. J Mol Biol 234:156–164

    Article  PubMed  CAS  Google Scholar 

  20. Essen LO, Siegert R, Lehmann WD, Oesterhelt D (1998) Lipid patches in membrane protein oligomers: crystal structure of the bacteriorhodopsin-lipid complex. Proc Natl Acad Sci USA 95:11673–11678

    Article  PubMed  CAS  Google Scholar 

  21. McDermott G, Prince SM, Freer AA, Hawthornthwaitelawless AM, Papiz MZ, Cogdell RJ, Isaacs NW (1995) Crystal structure of an integral membrane light-harvesting complex from photosynthetic bacteria. Nature 374:517–521

    Article  CAS  Google Scholar 

  22. Lancaster CRD, Kröger A, Auer M, Michel H (1999) Structure of fumarate reductase from Wolinella succinogenes at 2.2Å resolution. Nature 402:377–385

    Article  PubMed  CAS  Google Scholar 

  23. Hedderich T, Marcia M, Köpke J, Michel H (2011) PICKScreens, a new database for the comparison of crystallization screens for biological macromolecules. Cryst Growth Des 11:488–491

    Article  CAS  Google Scholar 

  24. Newby ZER, O’Connell JD, Gruswitz F, Hays FA, Harries WEC, Harwood IM, Ho JD, Lee JK, Savage DF, Miercke LJW, Stroud RM (2009) A general protocol for the crystallization of membrane proteins for X-ray structural investigation. Nat Protoc 4:619–637

    Article  PubMed  CAS  Google Scholar 

  25. Sumner JB, Dounce AL (1937) Crystalline catalase. Science 85:366–367

    Article  PubMed  CAS  Google Scholar 

  26. Wilkosz PA, Chandrasekhar K, Rosenberg JM (1995) Preliminary characterization of EcoRI*-DNA co-crystals—incomplete factorial design of oligonucleotide sequences. Acta Crystallogr D Biol Crystallogr 51: 938–945

    Article  PubMed  CAS  Google Scholar 

  27. IZIT User guide (http://hamptonresearch.com/documents/product/hr003025_4-710_izit_user_guide.pdf)

  28. Lebon G, Warne T, Tate CG (2012) Agonist-bound structures of G protein-coupled receptors. Curr Opin Struct Biol 22:482–490

    Article  PubMed  CAS  Google Scholar 

  29. Sciara G, Mancia F (2012) Highlights from recently determined structures of membrane proteins: a focus on channels and transporters. Curr Opin Struct Biol 22:476–481

    Article  PubMed  CAS  Google Scholar 

  30. PCT User Guide (http://hamptonresearch.com/documents/product/hr000257_2-140_user_guide.pdf)

  31. Newstead S, Ferrandon S, Iwata S (2008) Rationalizing alpha-helical membrane protein crystallization. Protein Sci 17:466–472

    Article  PubMed  CAS  Google Scholar 

  32. Parker JL, Newstead S (2012) Current trends in alpha-helical membrane protein crystallization: an update. Protein Sci 21:1358–1365

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Dr. Yvonne Carius for discussions and Saarland University, the Faculty of Medicine (HOMFOR), the state of Saarland (LFFP 11/02), and the Deutsche Forschungsgemeinschaft (DFG, grants INST 256/275-1 FUGG, GK 845-Lancaster, and GK1326-Lancaster) for supporting our research.

Author information

Authors and Affiliations

  1. Department of Structural Biology, Saarland University, Homburg, Germany

    Florian G. Müller & C. Roy D. Lancaster

  2. Center of Human and Molecular Biology (ZHMB), Saarland University, Homburg, Germany

    Florian G. Müller & C. Roy D. Lancaster

Authors
  1. Florian G. Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Roy D. Lancaster
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Interfaculty Institute of Biochemistry, University of Tuebingen, Tuebingen, Germany

    Doron Rapaport

  2. Cell Biology, University of Kaiserslautern, Kaiserslautern, Germany

    Johannes M. Herrmann

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Müller, F.G., Lancaster, C.R.D. (2013). Crystallization of Membrane Proteins. In: Rapaport, D., Herrmann, J. (eds) Membrane Biogenesis. Methods in Molecular Biology, vol 1033. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-487-6_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-487-6_5

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-486-9

  • Online ISBN: 978-1-62703-487-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation