Purification and crystallization of 15-lipoxygenase from rabbit reticulocytes.
We report a new purification of rabbit reticulocyte 15-lipoxygenase that has resulted in the first crystallization of a mammalian lipoxygenase. The enzyme was purified to homogeneity (greater than 98% pure by SDS-PAGE) using high pressure liquid chromatography on hydrophobic-interaction, hydroxyapatite and cation-exchange columns. Crystals were grown by the vapor diffusion method from concentrated solutions of the protein in sodium phosphate buffer, pH 7.0. The hexagonal, rod-shaped crystals were on average 0.09 mm x 0.09 mm x 0.4 mm, with approximate unit cell dimensions of a = b = 260 A, c = 145 A. The crystals diffract to 5 A resolution.
Sloane
D L
DL
Cardiovascular Research Institute (CVRI), University of California, San Francisco 94143.
Browner
M F
MF
Dauter
Z
Z
Wilson
K
K
Fletterick
R J
RJ
Sigal
E
E
D000818:Animals; D001093:Arachidonate 15-Lipoxygenase / Q000737:chemistry* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D004058:Diffusion; D004591:Electrophoresis, Polyacrylamide Gel; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology*; D014961:X-Ray Diffraction
D001093:Arachidonate 15-Lipoxygenase