Photosynthesis Research

, Volume 53, Issue 1, pp 35–44

Analysis of the chloroplast protein complexes by blue-native polyacrylamide gel electrophoresis (BN-PAGE)

  • Marion Kügler
  • Lothar Jänsch
  • Volker Kruft
  • Udo K. Schmitz
  • Hans-Peter Braun
Article

DOI: 10.1023/A:1005882406718

Cite this article as:
Kügler, M., Jänsch, L., Kruft, V. et al. Photosynthesis Research (1997) 53: 35. doi:10.1023/A:1005882406718

Abstract

Blue-native polyacrylamide gel electrophoresis (BN-PAGE) is a powerful procedure for the separation and characterization of the protein complexes from mitochondria. Membrane proteins are solubilized in the presence of aminocaproic acid and n-dodecylmaltoside and Coomassie-dyes are utilized before electrophoresis to introduce a charge shift on proteins. Here, we report a modification of the procedure for the analysis of chloroplast protein complexes. The two photosystems, the light-harvesting complexes, the ATP synthase, the cytochrome b6f complex and the ribulose-bisphosphate carboxylase/oxygenase are well resolved. Analysis of the protein complexes on a second gel dimension under denaturing conditions allows separation of more than 50 different proteins which are part of chloroplast multi-subunit enzymes. The resolution capacity of the blue-native gels is very high if compared to 'native green gel systems' published previously. N-terminal amino acid sequences of single subunits can be directly determined by cyclic Edman degradation as demonstrated for eight proteins. Analysis of chloroplast protein complexes by blue-native gel electrophoresis will allow the generation of 'protein maps' from different species, tissues and developmental stages or from mutant organelles. Further applications of blue-native gel electrophoresis are discussed.

b6f complex chloroplast ATP synthase light-harvesting complexes photosynthesis photosystems ribulose-bisphosphate carboxylase/oxygenase 

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Marion Kügler
    • 1
  • Lothar Jänsch
    • 1
  • Volker Kruft
    • 2
  • Udo K. Schmitz
    • 1
  • Hans-Peter Braun
    • 1
  1. 1.Institut für Angewandte GenetikUniversität HannoverHannoverGermany
  2. 2.Applied Biosystems GmbHWeiterstadtGermany

Personalised recommendations