Purification of His6-tagged Photosystem I from Chlamydomonas reinhardtii

Galina Gulis, Kuppala V. Narasimhulu, Lisa N. Fox, Kevin E. Redding

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

We have developed a rapid method for isolation of the Photosystem I (PS1) complex from Chlamydomonas reinhardtii using epitope tagging. Six histidine residues were genetically added to the N-terminus of the PsaA core subunit of PS1. The His6-tagged PS1 could be purified with a yield of 80-90% from detergent-solubilized thylakoid membranes within 3 h in a single step using a Ni-nitrilotriacetic acid (Ni-NTA) column. Immunoblots and low-temperature fluorescence analysis indicated that the His6-tagged PS1 preparation was highly pure and extremely low in uncoupled pigments. Moreover, the introduced tag appeared to have no adverse effect upon PS1 structure/function, as judged by photochemical assays and EPR spectroscopy of isolated particles, as well as photosynthetic growth tests of the tagged strain.

Original languageEnglish
Pages (from-to)51-60
Number of pages10
JournalPhotosynthesis Research
Volume96
Issue number1
DOIs
Publication statusPublished - Apr 1 2008

Keywords

  • Chlamydomonas reinhardtii
  • Epitope tagging
  • His-tag
  • Membrane protein
  • Photosystem 1
  • Reaction center

ASJC Scopus subject areas

  • Biochemistry
  • Plant Science
  • Cell Biology

Fingerprint Dive into the research topics of 'Purification of His<sub>6</sub>-tagged Photosystem I from Chlamydomonas reinhardtii'. Together they form a unique fingerprint.

  • Cite this