In vivo system for analyzing the function of the PsbP protein using Chlamydomonas reinhardtii
| Autor: | Fumihiko Sato, Taishi Nishimura, Kentaro Ifuku |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Photosystem II Protein subunit Mutant Chlamydomonas reinhardtii macromolecular substances Plant Science Photosynthesis Thylakoids Biochemistry Chromatography Affinity 03 medical and health sciences Botany Histidine Base Sequence biology Genetic Complementation Test Chlamydomonas Photosystem II Protein Complex food and beverages Cell Biology General Medicine biology.organism_classification Native Polyacrylamide Gel Electrophoresis Oxygen 030104 developmental biology Mutation Chromatography Gel Biophysics Green algae Peptides Oligopeptides Function (biology) |
| Zdroj: | Photosynthesis Research. 133:117-127 |
| ISSN: | 1573-5079 0166-8595 |
| DOI: | 10.1007/s11120-017-0370-2 |
| Popis: | The PsbP protein is an extrinsic subunit of photosystem II (PSII) specifically developed in green-plant species including land plants and green algae. The protein-protein interactions involving PsbP and its effect on oxygen evolution have been investigated in vitro using isolated PSII membranes. However, the importance of those interactions needs to be examined at the cellular level. To this end, we developed a system expressing exogenous PsbP in the background of the Chlamydomonas BF25 mutant lacking native PsbP. Expression of His-tagged PsbP successfully restored the oxygen-evolving activity and photoautotrophic growth of the mutant, while PsbP-∆15 lacking the N-terminal 15 residues, which are crucial for the oxygen-evolving activity of spinach PSII in vitro, only partially did. This demonstrated the importance of N-terminal sequence of PsbP for the photosynthetic activity in vivo. Furthermore, the PSII-LHCII supercomplex can be specifically purified from the Chlamydomonas cells having His-tagged PsbP using a metal affinity chromatography. This study provides a platform not only for the functional analysis of PsbP in vivo but also for structural analysis of the PSII-LHCII supercomplex from green algae. |
| Databáze: | OpenAIRE |
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