Expression, purification, crystallization and preliminary X-ray analysis of Pseudomonas aeruginosa AlgL.
The periplasmic alginate lyase AlgL is essential for the synthesis and export of the exopolysaccharide alginate in Pseudomonas sp. and also plays a role in its depolymerization. P. aeruginosa PAO1 AlgL has been overexpressed and purified and diffraction-quality crystals were grown using the hanging-drop vapour-diffusion method. The crystals grew as thin plates, with unit-cell parameters a = 56.4, b = 59.6, c = 102.1 Å, α = β = γ = 90°. The AlgL crystals exhibited the symmetry of space group P2(1)2(1)2(1) and diffracted to a minimum d-spacing of 1.64 Å. Based on the Matthews coefficient (V(M) = 2.20 Å(3) Da(-1)), one molecule is estimated to be present in the asymmetric unit.
Program in Molecular Structure and Function, Research Institute, Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada.
Francis
F
Wolfram
Kritica
K
Arora
Howard
H
Robinson
Ana Mirela
AM
Neculai
Patrick
P
Yip
P Lynne
PL
Howell
D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D011133:Polysaccharide-Lyases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011550:Pseudomonas aeruginosa / Q000201:enzymology*
D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase