Purification by dye-ligand chromatography and a crystallization study of the F1-ATPase and its major subunits, beta and alpha, from a thermophilic bacterium, PS3.
For a crystallization study, purification methods for F1-ATPase from a thermophilic bacterium, PS3, and its major subunits, beta and alpha, have been improved. The improvement depended on the introduction of dye-ligand chromatography columns to the previously adopted array of chromatography columns: a Blue-B (a blue dye bound to agarose) column was introduced for the F1 preparation, a Green-A column (a green dye attached to agarose) for the beta subunit, and a Blue-A (another blue dye, Cibacron Blue 3GA, bound to agarose) column for the alpha subunit. The improved preparations of all the proteins had purities of nearly 99%. Using the highly purified preparations of the proteins, crystallization conditions were searched for in a systematic way. Large plate crystals (0.2 X 0.5 X 0.5 mm) of F1 were grown from a polyethylene glycol solution. However, neither of the subunits was crystallized, in spite of extensive search for crystallization conditions.
Shirakihara
Y
Y
Department of Physics, Hyogo University of Education.
Yohda
M
M
Kagawa
Y
Y
Yokoyama
K
K
Yoshida
M
M
D001419:Bacteria / Q000201:enzymology*; D002845:Chromatography / Q000379:methods; D004396:Coloring Agents; D004926:Escherichia coli / Q000201:enzymology; D008024:Ligands; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000302:isolation & purification*; D012995:Solubility; D014961:X-Ray Diffraction
D004396:Coloring Agents; D008024:Ligands; D006180:Proton-Translocating ATPases