title: Isolation of hydrophobic membrane proteins by perfusion chromatography--purification of photosystem II reaction centers from spinach chloroplasts.
abstract: Ion exchange perfusion chromatography has been introduced for the isolation of hydrophobic membrane protein complexes from thylakoid membranes of spinach chloroplasts. By using this chromatographic technique, previously shown to be useful for the rapid isolation of soluble proteins (Regnier, F. E. (1991) Nature 350, 634-635), we have been able to isolate oxygen evolving photosystem II core complexes and photosystem II reaction center particles. Pure reaction centers could be isolated from photosystem II core complexes after a chromatographic step requiring only 6.5 mm, which is a substantial improvement in comparisons with previous procedures. The entire preparation of photosystem II core complexes and reaction center II particles could be completed in less than 2 h. The use of perfusion chromatography, as a versatile method for the isolation of hydrophobic membrane proteins from photosynthetic membranes, as well as for other biological membranes, will be discussed.
authors.affiliation: Department of Biochemistry, Stockholm University, Sweden.
authors.forename: M
authors.initials: M
authors.lastname: Roobol-Boza
authors.forename: B
authors.initials: B
authors.lastname: Andersson
mesh_terms: D002736:Chloroplasts / Q000737:chemistry*; D002845:Chromatography / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D003580:Cytochromes / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000302:isolation & purification*; D010455:Peptides / Q000032:analysis; D010477:Perfusion; D045322:Photosynthetic Reaction Center Complex Proteins / Q000302:isolation & purification*; D045332:Photosystem II Protein Complex; D010860:Pigments, Biological / Q000032:analysis; D013054:Spectrophotometry, Atomic; D018724:Spinacia oleracea
chemical_list: D003580:Cytochromes; D008565:Membrane Proteins; D010455:Peptides; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D010860:Pigments, Biological