Fast Isolation of Highly Active Photosystem II Core Complexes from Spinach
Zhao-Gai Wang1,2,3, Tian-Hua Xu1,2, Cheng Liu1 and Chun-Hong Yang1
1Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing, 100093, China 2Graduate University of Chinese Academy of Sciences, Beijing, 100049, China 3Institute of Farm Product Processing, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
Author for correspondence Tel: +86 10 6283 6252; E-mail: yangch@ibcas. ac.cn
Online on 26 May 2010 at www.jipb.net and www.interscience.wiley.com/journal/jipb 10.1111/j.1744-7909.2010.00971.x
Abstract
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 ZG, Xu TH, Liu C, Yang CH (2010) Fast isolation of highly active photosystem II core complexes from spinach. J. Integr. Plant Biol. 52(9), 793–800.
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Received 5 Jan 2010 Accepted 12 May 2010
© 2009 Institute of Botany, Chinese Academy of Sciences
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