Molecular Life Sciences

Living Edition
| Editors: Robert D. Wells, Judith S. Bond, Judith Klinman, Bettie Sue Siler Masters, Ellis Bell

Obtaining Crystals

  • Wasantha K. Ranatunga
  • Dan Su
  • Michael F. Romero
Living reference work entry
DOI: https://doi.org/10.1007/978-1-4614-6436-5_28-2

Synopsis

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...

Keywords

Protein Solution Quaternary Structure Vapor Diffusion Precipitant Concentration Crystallization Solution 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Notes

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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Wasantha K. Ranatunga
    • 1
  • Dan Su
    • 2
  • Michael F. Romero
    • 1
  1. 1.Department of Physiology and Biomedical Engineering & Nephrology & HypertensionMayo Clinic College of MedicineRochesterUSA
  2. 2.State Key Laboratory of BiotherapyWest China Hospital, Huaxi Campus, Sichuan UniversityChengduPeople’s Republic of China