Simultaneous refolding of denatured PsbS and reconstitution with LHCII into liposomes of thylakoid lipids

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• 作者：Cheng Liu ; Zhimin Gao ; Kun Liu ; Ruixue Sun ; Chunbo Cui…
• 关键词：Photosynthesis ; PsbS ; Proteoliposomes ; Reconstitution ; Refolding
• 刊名：Photosynthesis Research
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：127
• 期：1
• 页码：109-116
• 全文大小：1,331 KB
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• 作者单位：Cheng Liu (1)
  Zhimin Gao (2)
  Kun Liu (1)
  Ruixue Sun (1)
  Chunbo Cui (1)
  Alfred R. Holzwarth (3)
  Chunhong Yang (1)
  
  1. Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing, 100093, People鈥檚 Republic of China
  2. International Center for Bamboo and Rattan, State Forestry Administration Key Open Laboratory on Bamboo and Rattan Science and Technology, Beijing, 100102, People鈥檚 Republic of China
  3. Max-Planck-Institut f眉r Chemische Energiekonversion, Stiftstrasse 34-36, 45470, M眉lheim a. d. Ruhr, Germany
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Plant Physiology
  
• 出版者：Springer Netherlands
• ISSN：1573-5079

文摘

The thylakoid membrane protein PsbS is critical for quenching excessive excitation energy in mechanisms that involve the light-harvesting complexes of photosystem II. Liposomes of thylakoid lipids have been shown to be a very good platform to study photosynthetic membrane proteins and their interactions. In this study, we simultaneously refolded and reconstituted functional pea PsbS into liposomes of thylakoid lipids starting from denatured expressed protein. Intrinsic fluorescence spectroscopy, trypsin digestion, and circular dichroism spectroscopy were used to characterize the native state of PsbS in the proteoliposomes. The functionality of refolded PsbS was further demonstrated by its effect on the fluorescence quenching of the major antenna system of photosystem II (LHCII) co-inserted into the liposomes. The fluorescence yield of native trimeric LHCII was lowered by PsbS by 50 % at neutral pH and by a further 25 % upon lowering the pH to 4.5. Furthermore, the acid-induced fluorescence reduction was completely reversed by addition of N,N鈥?dicyclohexylcarbodiimide, an inhibitor of protein protonation. These results indicate that reconstituted PsbS induces strong quenching of LHCII sensing changes in local pH via its protonation sites. Keywords Photosynthesis PsbS Proteoliposomes Reconstitution Refolding