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New techniques for membrane protein crystallization tested on photosystem II core complex of Pisum sativum

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Abstract

The crystallization of a given protein is a hard task being even more complicated when the protein shows a hydrophobic behavior. In the case of photosynthetic proteins, the difficulty of the experiments increased due to the high light sensitivity. Aqueous solutions of photosystem II core complex (OEC PSII) of Pisum sativum were screened for crystallization conditions using standard crystallization methods. Crystal improvement was achieved by counter-diffusion technique in single capillaries of 0.2 mm inner diameter with a three-layer configuration. The use of this advanced crystallization technique—for the first time applied to the crystallization of membrane proteins—improves the reproducibility of the experiments allowing the initial crystal characterization, and facilitates the manipulation under light protection.

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Fig. 1
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Abbreviations

OEC PSII:

Oxygen-evolving complex of photosystem II or photosystem II core complex

PSII:

Photosystem II

MPDB:

Membrane protein data bank

Fe :

Iron

Mn :

Manganese

Chla :

Chlorophyll a

PEG4000 :

Polyethylenglycol Mr 4000

Bis–Tris :

Bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane

MES :

2-(N-morpholino)ethanesulfonic acid

MPD :

2-Methyl-2,4-pentanediol

DM:

n-dodecyl-β-D-maltoside

References

  • Adir N (1999) Crystallization of the oxygen-evolving reaction centre of photosystem II in nine different detergent mixtures. Acta Cryst D55:891–894

    CAS  Google Scholar 

  • Barber J (2003) Photosystem II: the engine of life. Q Rev Biophys 36:71–89

    Article  PubMed  CAS  Google Scholar 

  • Bergfors TM (ed) (1999) Protein Crystallization: techniques, strategies, and tips. International University Line, La Jolla

  • Ducruix A, Giege R (eds) (1999) Crystallization of nucleic acids and proteins. Oxford University Press, Oxford

  • Ferreira KN, Iverson TM, Maghlaoui K, Barber J, Iwata S (2004) Architecture of the photosynthetic oxygen-evolving center. Science 303:1831–1838

    Article  PubMed  CAS  Google Scholar 

  • Garcia-Ruiz JM (2003) Counter-diffusion methods for macromolecular crystallization. Methods in Enzymology, vol 368. Academic Press, pp 130–154

  • Gavira JA, de Jesus W, Camara-Artigas A, Lopez-Garriga J, Garcia-Ruiz JM (2006) Capillary crystallization and molecular-replacement solution of haemoglobin II from the clam Lucina pectinata. Acta Cryst F62:196–199

    CAS  Google Scholar 

  • Ghanotakis DF, Demetriou DM, Yocum CF (1987) Isolation and characterization of an oxygen-evolving photosystem-II reaction center core preparation and a 28 kDa chl-alpha-binding protein. Biochim Biophys Acta 891:15–21

    Article  CAS  Google Scholar 

  • Kamiya N, Shen JR (2003) Crystal structure of oxygen-evolving photosystem II from Thermosynechococcus vulcanus at 3.7 Å resolution. PNAS 100:98–103

    Article  PubMed  CAS  Google Scholar 

  • Kern J, Loll B, Lueneberg C, DiFiore D, Biesiadka J, Irrgang K-D, Zouni A (2005) Purification, characterisation and crystallization of photosystem II from Thermosynechococcus elongatus cultivated in a new type of photobioreactor. BBA 1706:147–157

    Article  PubMed  CAS  Google Scholar 

  • Kuhl H, Kruip J, Seidler A, Krieger-Liszkay A, Bünker M, Bald D, Scheidig AJ, Rögner M (2000) Towards structural determination of the water-splitting enzyme. J Biol Chem 275:20652–20659

    Article  PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Loll B, Kern J, Saenger W, Zouni A, Biesiadka J (2005) Towards complete cofactor arrangement in the 3.0 Å resolution structure of photosystem II. Nat Lett 438:1040–1044

    Article  CAS  Google Scholar 

  • Lopez-Jaramillo FJ, Garcia-Ruiz JM, Gavira JA, Otalora F (2001) Crystallization and cryocrystallography inside X-ray capillaries. J Appl Cryst 34:365–370

    Article  CAS  Google Scholar 

  • Raman P, Cherezov V, Caffrey M (2006) The membrane protein data bank. Cell Mol Life Sci 63:36–51, http://www.mpdb.ul.ie/index.asp

    Google Scholar 

  • Shen J-R, Kamiya N (2000) Crystallization and the crystal properties of the oxygen-evolving photosystem II from Synechococcus vulcanus. Biochemistry 39:14739–14744

    Article  PubMed  CAS  Google Scholar 

  • Zouni A, Witt H-T, Kern J, Fromme P, Krauss N, Saenger W, Orth P (2001) Crystal structure of photosystem II from Synechococcus elongates at 3.8 Å resolution. Nature 409:739–743

    Article  PubMed  CAS  Google Scholar 

  • Wydrzynski T, Satoh K (eds) (2005) Photosystem II, the light driven water: plastoquinone oxidoreductase. Springer, Dordrecht

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Acknowledgments

This work is supported by the Grant Agency of the Czech Republic (206/03/D061), by the joint CSIC and AS CR projects 2003CZ0013 and 2004CZ0003, by the Ministry of Education of the Czech Republic (MSM6007665808, LC06010) and by the Academy of Sciences of the Czech Republic (AV0Z60870520, AV0Z50510513). We also acknowledge Junta de Andalucía (Spain) for financial support through the project Excelencia (RNM1344) and support to JAG. IKS thanks to Marie Šáchová and Ivana Hunalová for protein sample preparation. We are grateful to X13, BM16 and ID14-1 staff for the support during data collection.

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Author notes

  1. Pavlína Řezáčová

    Present address: Department of Biochemistry, UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TA, 75390-8816, USA

Authors and Affiliations

  1. Institute of Physical Biology, University of South Bohemia Ceske Budejovice, Zamek 136, 373 33, Nove Hrady, Czech Republic

    Ivana Kutá Smatanová & František Vácha

  2. Institute of Systems Biology and Ecology AS CR, Zamek 136, 373 33, Nove Hrady, Czech Republic

    Ivana Kutá Smatanová

  3. Laboratorio de Estudios Cristalografico, Edf. Lopez Neira, P.T. Ciencias de la Salud, Avenida del Conocimiento, s/n, 18100, Armilla, Granada, Spain

    José A. Gavira & Juan M. García-Ruiz

  4. Institute of Molecular Genetics AS CR, Flemingovo n. 2, 16637, Prague, Czech Republic

    Pavlína Řezáčová

  5. Biological Centre IPMB AS CR, Branisovska 31, 370 05, Ceske Budejovice, Czech Republic

    František Vácha

Authors
  1. Ivana Kutá Smatanová
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  2. José A. Gavira
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  3. Pavlína Řezáčová
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  4. František Vácha
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  5. Juan M. García-Ruiz
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Correspondence to Ivana Kutá Smatanová.

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Kutá Smatanová, I., Gavira, J.A., Řezáčová, P. et al. New techniques for membrane protein crystallization tested on photosystem II core complex of Pisum sativum . Photosynth Res 90, 255–259 (2006). https://doi.org/10.1007/s11120-007-9131-y

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  • DOI: https://doi.org/10.1007/s11120-007-9131-y

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