Biochemical and structural studies of the large ycf4-photosystem i assembly complex of the green alga chlamydomonas reinhardtii

Shinichiro Ozawa, Jon Nield, Akihiro Terao, Einar J. Stauber, Michael Hippler, Hiroyuki Koike, Jean David Rochaix, Yuichiro Takahashi

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Ycf4 is a thylakoid protein essential for the accumulation of photosystem I (PSI) in Chlamydomonas reinhardtii. Here, a tandem affinity purification tagged Ycf4 was used to purify a stable Ycf4-containing complex of >1500 kD. This complex also contained the opsin-related COP2 and the PSI subunits PsaA, PsaB, PsaC, PsaD, PsaE, and PsaF, as identified by mass spectrometry (liquid chromatography-tandem mass spectrometry) and immunoblotting. Almost all Ycf4 and COP2 in wildtype cells copurified by sucrose gradient ultracentrifugation and subsequent ion exchange column chromatography, indicating the intimate and exclusive association of Ycf4 and COP2. Electron microscopy revealed that the largest structures in the purified preparation measure 285 x 185 Á these particles may represent several large oligomeric states. Pulse-chase protein labeling revealed that the PSI polypeptides associated with the Ycf4-containing complex are newly synthesized and partially assembled as a pigment-containing subcomplex. These results indicate that the Ycf4 complex may act as a scaffold for PSI assembly. A decrease in COP2 to 10% of wild-type levels by RNA interference increased the salt sensitivity of the Ycf4 complex stability but did not affect the accumulation of PSI, suggesting that COP2 is not essential for PSI assembly.

Original languageEnglish
Pages (from-to)2424-2442
Number of pages19
JournalPlant Cell
Volume21
Issue number8
DOIs
Publication statusPublished - Aug 2009

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Photosystem I Protein Complex
Chlamydomonas reinhardtii
Chlorophyta
photosystem I
Opsins
Thylakoids
Ultracentrifugation
Ion Exchange Chromatography
opsin
Tandem Mass Spectrometry
RNA Interference
ultracentrifugation
Immunoblotting
Liquid Chromatography
ion exchange
Sucrose
immunoblotting
Mass Spectrometry
RNA interference
Electron Microscopy

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology

Cite this

Biochemical and structural studies of the large ycf4-photosystem i assembly complex of the green alga chlamydomonas reinhardtii. / Ozawa, Shinichiro; Nield, Jon; Terao, Akihiro; Stauber, Einar J.; Hippler, Michael; Koike, Hiroyuki; Rochaix, Jean David; Takahashi, Yuichiro.

In: Plant Cell, Vol. 21, No. 8, 08.2009, p. 2424-2442.

Research output: Contribution to journalArticle

Ozawa, Shinichiro ; Nield, Jon ; Terao, Akihiro ; Stauber, Einar J. ; Hippler, Michael ; Koike, Hiroyuki ; Rochaix, Jean David ; Takahashi, Yuichiro. / Biochemical and structural studies of the large ycf4-photosystem i assembly complex of the green alga chlamydomonas reinhardtii. In: Plant Cell. 2009 ; Vol. 21, No. 8. pp. 2424-2442.
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