title: Detergent effect on anion exchange perfusion chromatography and gel filtration of intact chloroplast H(+)-ATP synthase.
abstract: To gain a pure enzyme preparation for functional and crystallization studies, an additional purification step in the isolation of the chloroplast ATP synthase (CF(0)F(1)) has been introduced. By applying gel filtration or anion exchange perfusion chromatography in presence of the detergents CHAPS and n-dodecyl-beta-d-maltoside, respectively, Rubisco and other contaminants were separated from CF(0)F(1). The purity and activity depended on the chromatographic method and the detergent employed. The highest purity and activity were achieved by anion exchange chromatography for the detergent dodecyl-maltoside and by gel filtration for the detergent CHAPS. The detergent Triton X-100, which is frequently used to solubilize CF(0)F(1), was found to be inadequate to stabilize the ATP synthase during chromatography.
authors.affiliation: Abteilung Physikalische Biochemie, Technische Universität Darmstadt, Petersenstrasse 22, Darmstadt, D-64287, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Poetsch
authors.forename: H
authors.initials: H
authors.lastname: Seelert
authors.forename: J
authors.initials: J
authors.lastname: Meyer zu Tittingdorf
authors.forename: N A
authors.initials: NA
authors.lastname: Dencher
mesh_terms: D002736:Chloroplasts / Q000201:enzymology*; D002793:Cholic Acids; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003902:Detergents; D004795:Enzyme Stability; D005069:Evaluation Studies as Topic; D005960:Glucosides; D017830:Octoxynol; D006180:Proton-Translocating ATPases / Q000302:isolation & purification*; D012995:Solubility; D018724:Spinacia oleracea / Q000201:enzymology
chemical_list: D002793:Cholic Acids; D003902:Detergents; D005960:Glucosides; C040358:dodecyl maltoside; D017830:Octoxynol; D006180:Proton-Translocating ATPases; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate