Detergent effect on anion exchange perfusion chromatography and gel filtration of intact chloroplast H+-ATP synthase

To gain a pure enzyme preparation for functional and crystallization studie s, an additional purification step in the isolation of the chloroplast ATP synthase (CF0F1) has been introduced. By applying gel filtration or anion e xchange perfusion chromatography in presence of the detergents CHAPS and n- dodecyl-beta-D-maltoside, respectively, Rubisco and other contaminants were separated from CF0F1. The purity and activity depended on the chromatograp hic method and the detergent employed, The highest purity and activity were achieved by anion exchange chromatography for the detergent dodecyl-maltos ide and by gel filtration for the detergent CHAPS. The detergent Triton X-1 00, which is frequently used to solubilize CF0F1, was found to be inadequat e to stabilize the ATP synthase during chromatography, (C) 1999 Academic Pr ess.