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Obtaining Crystals

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Protein structure determination is one of the important steps for understanding the function of biological systems. Purified protein solution is used for setting up crystal screens. Obtaining pure protein solution generally involves two or more purification steps, and optimization of each step is needed. Better expression systems should be explored for each protein of interest for obtaining reasonable amount of proteins (2–5 mg) for crystallization. Purified protein has to be stable, homogenous, and monodisperse for the fulfillment of crystallization. Protein crystallization is performed by mixing protein solution with different conditions (buffers, additives, and precipitants) at microliter scale (0.1–20 μL) in a tray and incubated for a period of time. Different methods and screenings for setting up crystal trays can be explored for each protein. The most common method is called vapor diffusion with hanging drop and sitting drop methods. More automated methods of...

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References

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

    Google Scholar 

  • Bergfors T (2003) Seeds to crystals. J Struct Biol 142:66–76

    Article  CAS  PubMed  Google Scholar 

  • Chayen NE, Saridakis E (2008) Protein crystallization: from purified protein to diffraction-quality crystal. Nat Methods 5:147–153

    Article  CAS  PubMed  Google Scholar 

  • Hassell AM, An G, Bledsoe RK, Bynum JM, Carter HL III, Deng SJ, Gampe RT, Grisard TE, Madauss KP, Nolte RT, Rocque WJ, Wang L, Weaver KL, Williams SP, Wisely GB, Xu R, Shewchuk LM (2007) Crystallization of protein-ligand complexes. Acta Cryst D63:72–79

    Google Scholar 

  • Saridakis E, Chayen NE (2008) Towards a ‘universal’ nucleant for protein crystallization. Cell 27:99–106

    Google Scholar 

  • Wiener MC (2004) A pedestrian guide to membrane protein crystallization. Methods 34:364–372

    Article  CAS  PubMed  Google Scholar 

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Acknowledgement

The authors would like to thank Dr. James Thompson and the members of the Romero laboratory for thoughtful comments on the manuscript. This work was supported by NIH grants EY017732 & DK092408 (MFR) and the Mayo Foundation.

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Correspondence to Wasantha K. Ranatunga .

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© 2014 Springer Science+Business Media New York

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Ranatunga, W.K., Su, D., Romero, M.F. (2014). Obtaining Crystals. In: Bell, E. (eds) Molecular Life Sciences. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-6436-5_28-2

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  • DOI: https://doi.org/10.1007/978-1-4614-6436-5_28-2

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  • Publisher Name: Springer, New York, NY

  • Online ISBN: 978-1-4614-6436-5

  • eBook Packages: Springer Reference Biomedicine and Life SciencesReference Module Biomedical and Life Sciences

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