Novel, rapid purification of the membrane protein photosystem I by high-performance liquid chromatography on porous materials.
New porous materials have been tested for their potential to speed up purification of membrane proteins. As an example the purification of photosystem I, a light-driven electron pump from the cyanobacterium Synechocystis PCC6803, was optimized. The combination of two HPLC steps (an anion-exchange chromatography followed by a hydrophobic interaction chromatography) yields homogeneous monomeric or trimeric photosystem I as determined by gel filtration and gel electrophoresis. In comparison to traditional purification schemes our method is at least three-times faster and allows for easy scale-up.
Wenk
S O
SO
University of Bochum, Plant Biochemistry, Faculty of Biology, Germany.
Kruip
J
J
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods; D000458:Cyanobacteria / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000302:isolation & purification*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000302:isolation & purification*
D008565:Membrane Proteins; D045322:Photosynthetic Reaction Center Complex Proteins