title: Crystallization of Pseudomonas aeruginosa AmrZ protein: development of a comprehensive method for obtaining and optimization of protein-DNA crystals.
abstract: The AmrZ protein from the pathogenic bacterium Pseudomonas aeruginosa is a transcription factor that activates and represses the genes for several potent virulence factors, which gives the bacteria a selective advantage in infection. AmrZ was crystallized in complex with DNA containing the amrZ1 repressor binding site. Obtaining crystals of the complex required the integration of a number of well known techniques along with the development of new methods. Here, these processes are organized and combined into a comprehensive method which yielded diffraction-quality crystals. Part of this method included thorough data mining of the crystallization conditions of protein-DNA complexes to create a new directed crystallization screen. An optimized technique for the verification of protein-DNA complexes in crystals is also presented. Taken together, the methods described in this article attempt to streamline the difficult process of obtaining diffraction-quality crystals of protein-DNA complexes through the organization of older methods combined with the introduction of new techniques.
authors.affiliation: Department of Biochemistry and Center for Structural Biology, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
authors.forename: Edward E
authors.initials: EE
authors.lastname: Pryor
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Wozniak
authors.forename: Thomas
authors.initials: T
authors.lastname: Hollis
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry*; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray; D004247:DNA / Q000032:analysis / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011550:Pseudomonas aeruginosa / Q000737:chemistry*; D014157:Transcription Factors / Q000032:analysis / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D014157:Transcription Factors; D004247:DNA