title: Crystallization, dehydration and preliminary X-ray analysis of excisionase (Xis) proteins cooperatively bound to DNA.
abstract: This paper describes the crystallization, dehydration and preliminary X-ray data analysis of a complex containing several bacteriophage lambda excisionase (Xis) [Bushman et al. (1984). Cell, 39, 699-706] proteins cooperatively bound to a 33-mer DNA duplex (Xis-DNA(X1-X2)). Xis is expected to recognize this regulatory element in a novel manner by cooperatively binding and distorting multiple head-to-tail orientated DNA-binding sites. Crystals of this complex belonged to space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 107.7, c = 73.5 angstroms, alpha = beta = 90, gamma = 120 degrees. Based on the unit-cell parameters for the asymmetric unit, V(M) is 3.0 A(3) Da(-1), which corresponds to a solvent content of approximately 59%. The approaches used to crystallize the unusually long DNA fragment in the complex and the dehydration technique applied that dramatically improved the diffraction of the crystals from 10 to 2.6 angstroms are discussed.
authors.affiliation: Department of Chemistry and Biochemistry and the UCLA-DOE Center for Genomics and Proteomics, University of California, Los Angeles, 405 Hilgard Avenue, Los Angeles, CA 90095-1570, USA.
authors.forename: My D
authors.initials: MD
authors.lastname: Sam
authors.forename: Mohamad A
authors.initials: MA
authors.lastname: Abbani
authors.forename: Duilio
authors.initials: D
authors.lastname: Cascio
authors.forename: Reid C
authors.initials: RC
authors.lastname: Johnson
authors.forename: Robert T
authors.initials: RT
authors.lastname: Clubb
mesh_terms: D010582:Bacteriophage lambda / Q000201:enzymology; D001483:Base Sequence; D001665:Binding Sites; D003460:Crystallization; D004254:DNA Nucleotidyltransferases / Q000737:chemistry* / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D011485:Protein Binding; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D014764:Viral Proteins; D004254:DNA Nucleotidyltransferases; C063911:excisionase