Purification of Multisubunit Membrane Protein Complexes: Isolation of Chloroplast FoF1-ATP Synthase, CFo and CF1 by Blue Native Electrophoresis

1999 ◽  
Vol 259 (3) ◽  
pp. 569-575 ◽  
Author(s):  
Dirk Neff ◽  
Norbert A. Dencher
Keyword(s):  
Membrane Protein ◽  
Atp Synthase ◽  
Protein Complexes ◽  
Native Electrophoresis ◽  
Membrane Protein Complexes ◽  
Blue Native Electrophoresis ◽  
Blue Native ◽  
Fof1 Atp Synthase

LC-nanospray-MS/MS analysis of hydrophobic proteins from membrane protein complexes isolated by blue-native electrophoresis

2005 ◽  
Vol 40 (9) ◽  
pp. 1223-1231 ◽  
Author(s):  
Anabel S. Fandiño ◽  
Isam Rais ◽  
Martin Vollmer ◽  
Helmuth Elgass ◽  
Hermann Schägger ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Complexes ◽  
Ms Analysis ◽  
Native Electrophoresis ◽  
Membrane Protein Complexes ◽  
Blue Native Electrophoresis ◽  
Blue Native

scholarly journals Dye-ligand chromatographic purification of intact multisubunit membrane protein complexes: application to the chloroplast H+-F0F1-ATP synthase

2000 ◽  
Vol 346 (1) ◽  
pp. 41-44
Author(s):  
Holger SEELERT ◽  
Ansgar POETSCH ◽  
Meino ROHLFS ◽  
Norbert A. DENCHER
Keyword(s):  
Membrane Protein ◽  
Phosphatidic Acid ◽  
Ammonium Sulphate ◽  
Atp Synthase ◽  
Protein Complex ◽  
Protein Complexes ◽  
Atp Synthesis ◽  
Chromatographic Purification ◽  
Membrane Protein Complexes ◽  
Fof1 Atp Synthase

n-Dodecyl-β-D-maltoside was used as a detergent to solubilize the ammonium sulphate precipitate of chloroplast FOF1-ATP synthase, which was purified further by dye-ligand chromatography. Upon reconstitution of the purified protein complex into phosphatidylcholine/phosphatidic acid liposomes, ATP synthesis, driven by an artificial ∆pH/∆ψ, was observed. The highest activity was achieved with ATP synthase solubilized in n-dodecyl-β-D-maltoside followed by chromatography with Red 120 dye. The optimal dye for purification with CHAPS was Green 5. All known subunits were present in the monodisperse proton-translocating ATP synthase preparation obtained from chloroplasts.

Blue native electrophoresis as a tool for detection of PIP-containing protein complexes in the plasma membranes

2013 ◽  
Vol 60 (4) ◽  
pp. 563-570
Author(s):  
T. A. Shevyreva ◽  
M. S. Piotrovskii ◽  
B. V. Belugin ◽  
I. M. Zhestkova ◽  
M. S. Trofimova
Keyword(s):  
Plasma Membranes ◽  
Protein Complexes ◽  
Native Electrophoresis ◽  
Blue Native Electrophoresis ◽  
Blue Native

scholarly journals A Novel Chloroplast Super-Complex Consisting of the ATP Synthase and Photosystem I Reaction Center

2019 ◽  
Author(s):  
Satarupa Bhaduri ◽  
Sandeep K Singh ◽  
Whitaker Cohn ◽  
S. Saif Hasan ◽  
Julian P. Whitelegge ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Membrane Protein ◽  
Reaction Center ◽  
Atp Synthase ◽  
Photosystem I ◽  
Protein Complexes ◽  
Thylakoid Membranes ◽  
Native Page ◽  
Membrane Protein Complexes ◽  
Reaction Center Complexes

AbstractSeveral ‘super-complexes’ of individual hetero-oligomeric membrane protein complexes, whose function is to facilitate intra-membrane electron and proton transfer and harvesting of light energy, have been previously characterized in the mitochondrial cristae and chloroplast thylakoid membranes. The latter membrane is reported here to also be the location of an intra-membrane super-complex which is dominated by the ATP-synthase and photosystem I (PSI) reaction-center complexes, defined by mass spectrometry, clear-native PAGE and Western Blot analyses. This is the first documented presence of ATP synthase in a super-complex with the PSI reaction-center located in the non-appressed stromal domain of the thylakoid membrane.

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