Fast isolation of highly active photosystem II core complexes from spinach.
Purification of photosystem II (PSII) core complexes is a time-consuming and low-efficiency process. In order to isolate pure and active PSII core complexes in large amounts, we have developed a fast method to isolate highly active monomeric and dimeric PSII core complexes from spinach leaves by using sucrose gradient ultracentrifugation. By using a vertical rotor the process was completed significantly faster compared with a swing-out rotor. In order to keep the core complexes in high activity, the whole isolation procedure was performed in the presence of glycine betain and pH at 6.3. The isolated pigment-protein complexes were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, absorption spectroscopy, 77 K fluorescence spectroscopy and high performance liquid chromatography. Our results show that this method is a better choice for quick and efficient isolation of functionally active PSII core complexes.
Wang
Zhao-Gai
ZG
Key Laboratory of Photobiology, Institute of Botany, the Chinese Academy of Sciences, Beijing, China.
Xu
Tian-Hua
TH
Liu
Cheng
C
Yang
Chun-Hong
CH
D002851:Chromatography, High Pressure Liquid; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D045332:Photosystem II Protein Complex / Q000302:isolation & purification*; D013050:Spectrometry, Fluorescence; D018724:Spinacia oleracea / Q000737:chemistry*; D014461:Ultracentrifugation
D045332:Photosystem II Protein Complex