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Combining Wet and Dry Lab Techniques to Guide the Crystallization of Large Coiled-coil Containing Proteins
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Biochemistry
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JoVE Journal Biochemistry
Combining Wet and Dry Lab Techniques to Guide the Crystallization of Large Coiled-coil Containing Proteins

Combining Wet and Dry Lab Techniques to Guide the Crystallization of Large Coiled-coil Containing Proteins

DOI:
10.3791/54886-v

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11:14 min

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January 06, 2017

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, , , , ,
1Department of Biological Sciences,University of Pittsburgh, 2Institute of Structural Biology,German Research Center for Environmental Health

Chapters

  • 00:05Title
  • 00:48Express and Purify Proteins with Identified Domain Boundaries
  • 02:38Characterize Protein Sample to Identify Those with Advantageous Properties
  • 04:16Producing High Quality Crystals of the Coiled-coil SD2 Domain from Shroom
  • 08:43Results: Evaluation of Expression, Purification, Characterization, and Crystallization of Candidate Coiled-coil Proteins
  • 09:49Conclusion

Summary

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Automatic Translation

We describe a framework incorporating straightforward biochemical and computational analysis to guide the characterization and crystallization of large coiled-coil domains. This framework can be adapted for globular proteins or extended to incorporate a variety of high-throughput techniques.

Tags

Keywords: Coiled-coil ProteinsCrystallizationProtein PurificationNickel Affinity PurificationSDS-PAGENative PAGEUV-Vis Spectroscopy

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