Photosynthesis Research

, Volume 96, Issue 1, pp 51–60

Purification of His6-tagged Photosystem I from Chlamydomonas reinhardtii

  • Galina Gulis
  • Kuppala V. Narasimhulu
  • Lisa N. Fox
  • Kevin E. Redding
Regular Paper

DOI: 10.1007/s11120-007-9283-9

Cite this article as:
Gulis, G., Narasimhulu, K.V., Fox, L.N. et al. Photosynth Res (2008) 96: 51. doi:10.1007/s11120-007-9283-9

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.

Keywords

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

Supplementary material

11120_2007_9283_Fig9_ESM.jpg (178 kb)
Supplementary Fig. 1(JPG 177 kb)

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Galina Gulis
    • 1
  • Kuppala V. Narasimhulu
    • 1
  • Lisa N. Fox
    • 1
  • Kevin E. Redding
    • 1
    • 2
  1. 1.Departments of Chemistry and Biological SciencesUniversity of AlabamaTuscaloosaUSA
  2. 2.Institute de Biologie Physico-ChimiqueParisFrance

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