Novel, rapid purification of the membrane protein photosystem I by high-performance liquid chromatography on porous materials

• Published in: Journal of chromatography. B, Biomedical sciences and applications Vol. 737; no. 1-2; pp. 131 - 142
• Main Authors: Wenk, Stephan-Olav, Kruip, Jochen
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 14.01.2000; Elsevier Science
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Chromatography, Gel; Chromatography, High Pressure Liquid - methods; Chromatography, Ion Exchange - methods; Cyanobacteria - chemistry; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; Hydrogen-Ion Concentration; Membrane proteins; Membrane Proteins - isolation & purification; Microbiology; Photosynthetic Reaction Center Complex Proteins - isolation & purification; Photosystem I; Proteins; Proteins; Photosystem I; Synechocystis; Purification; Membrane proteins; Photosystem 1; Separation method; Membrane protein; Cyanobacteria; HPLC chromatography; Bacteria
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(99)00525-3

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.