Expression, purification, crystallization and preliminary X-ray analysis of Pseudomonas fluorescens AlgK.
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.
Keiski
Carrie-Lynn
CL
Program in Molecular Structure and Function, Research Institute, Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada.
Yip
Patrick
P
Robinson
Howard
H
Burrows
Lori L
LL
Howell
P Lynne
PL
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*
C104798:AlgK protein, Pseudomonas aeruginosa; D001426:Bacterial Proteins; D017931:DNA Primers