Crystallization of protein-DNA complexes.
Determining the crystal structure of a protein-DNA complex can provide a wealth of information regarding protein function and mechanism. The foundation for all successful X-ray structure determination is the ability to produce diffraction-quality crystals. Crystallization of protein-DNA complexes often presents unique challenges because of the additional parameters involved. This chapter will outline many of those challenges, including choice of DNA and formation of a stable protein-DNA complex and provide guidance in preparing for crystallization experiments. Additionally, techniques for oligonucleotide purification, sample preparation, and crystallization methods are provided. Careful thought and initial analysis of the protein-DNA complex prior to crystallization experiments followed by optimization of crystallization parameters can greatly increase the likelihood of producing well-diffracting crystals.
Hollis
Thomas
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Department of Biochemistry, Center for Structural Biology, Wake Forest University Health Science Center, Winston Salem, NC, USA.
D003460:Crystallization / Q000379:methods*; D004247:DNA / Q000737:chemistry; D046911:Macromolecular Substances / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry
D046911:Macromolecular Substances; D011506:Proteins; D004247:DNA