Apr, 1998
Rapid and discrete isolation of oxygen-evolving His-tagged photosystem II core complex from Chlamydomonas reinhardtii by Ni2+ affinity column chromatography
FEBS LETTERS
- ,
- ,
- Volume
- 426
- Number
- 1
- First page
- 140
- Last page
- 144
- Language
- English
- Publishing type
- Research paper (scientific journal)
- DOI
- 10.1016/S0014-5793(98)00328-7
- Publisher
- ELSEVIER SCIENCE BV
We have developed a simple and rapid procedure to isolate an oxygen-evolving photosystem II (PS II) core complex from Chlamydomonas reinhardtii. A His-tag made of sis consecutive histidine residues was genetically attached at the carboxy terminus of D2 protein to create a metal binding site on the PS Il supramolecular complex, The recombinant cells producing the His-tagged variant of D2 protein grew photo-autotrophically as well as the wild-type cells. Characterization of the oxygen evolution and the thermoluminescence properties revealed that the His-tagging did not affect the functional integrity of the PS II reaction center. A PS Il core complex was isolated from the detergent-solubilized thylakoids of the recombinant cells in 4 h by a single one-step Ni2+ affinity column chromatography. This preparation consists of D1, D2, CP43, CP47, 33 kDa, and a few low molecular weight proteins, and retains a high rate of oxygen-evolving activity (=1000 mu mol/mg Chl/h), (C) 1998 Federation of European Biochemical Societies.
- Link information
-
- DOI
- https://doi.org/10.1016/S0014-5793(98)00328-7
- CiNii Articles
- http://ci.nii.ac.jp/naid/80010262064
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/9598995
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000073250000029&DestApp=WOS_CPL
- ID information
-
- DOI : 10.1016/S0014-5793(98)00328-7
- ISSN : 0014-5793
- CiNii Articles ID : 80010262064
- Pubmed ID : 9598995
- Web of Science ID : WOS:000073250000029