Rapid and discrete isolation of oxygen-evolving His-tagged photosystem II core complex from Chlamydomonas reinhardtii by Ni2+ affinity column chromatography.
FEBS Lett
; 426(1): 140-4, 1998 Apr 10.
Article
in En
| MEDLINE
| ID: mdl-9598995
ABSTRACT
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 six consecutive histidine residues was genetically attached at the carboxy terminus of D2 protein to create a metal binding site on the PS II supramolecular complex. The recombinant cells producing the His-tagged variant of D2 protein grew photoautotrophically 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 II 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 micromol/mg Chl/h).
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Chlamydomonas reinhardtii
/
Photosynthetic Reaction Center Complex Proteins
Limits:
Animals
Language:
En
Journal:
FEBS Lett
Year:
1998
Document type:
Article
Affiliation country:
Japan