title: Expression, purification, crystallization and preliminary X-ray analysis of Pseudomonas fluorescens AlgK.
abstract: AlgK is an outer-membrane lipoprotein involved in the biosynthesis of alginate in Pseudomonads and Azotobacter vinelandii. A recombinant form of Pseudomonas fluorescens AlgK with a C-terminal polyhistidine affinity tag has been expressed and purified from the periplasm of Escherichia coli cells and diffraction-quality crystals of AlgK have been grown using the hanging-drop vapour-diffusion method. The crystals grow as flat plates with unit-cell parameters a = 79.09, b = 107.85, c = 119.15 A, beta = 96.97 degrees. The crystals exhibit the symmetry of space group P2(1) and diffract to a minimum d-spacing of 2.5 A at Station X29 of the National Synchrotron Light Source, Brookhaven National Laboratory. On the basis of the Matthews coefficient (V(M) = 2.53 A3 Da(-1)), four protein molecules are estimated to be present in the asymmetric unit.
authors.affiliation: Program in Molecular Structure and Function, Research Institute, Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada.
authors.forename: Carrie-Lynn
authors.initials: CL
authors.lastname: Keiski
authors.forename: Patrick
authors.initials: P
authors.lastname: Yip
authors.forename: Howard
authors.initials: H
authors.lastname: Robinson
authors.forename: Lori L
authors.initials: LL
authors.lastname: Burrows
authors.forename: P Lynne
authors.initials: PL
authors.lastname: Howell
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D011487:Protein Conformation; D011551:Pseudomonas fluorescens / Q000737:chemistry*
chemical_list: C104798:AlgK protein, Pseudomonas aeruginosa; D001426:Bacterial Proteins; D017931:DNA Primers