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.

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.

scholarly journals Surfactant-free purification of membrane protein complexes from bacteria: application to the staphylococcal penicillin-binding protein complex PBP2/PBP2a

2014 ◽  
Vol 25 (28) ◽  
pp. 285101 ◽  
Author(s):  
Sarah Paulin ◽  
Mohammed Jamshad ◽  
Timothy R Dafforn ◽  
Jorge Garcia-Lara ◽  
Simon J Foster ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Complex ◽  
Protein Complexes ◽  
Binding Protein ◽  
Penicillin Binding Protein ◽  
Membrane Protein Complexes

scholarly journals A Genetically Encoded Trimethylsilyl 1D 1H-NMR Probe for Conformation Change in Large Membrane Protein Complexes

2019 ◽  
Author(s):  
Qi Liu ◽  
Qing-tao He ◽  
Xiao-xuan Lyu ◽  
Fan Yang ◽  
Zhong-liang Zhu ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Complex ◽  
Protein Complexes ◽  
Signal To Noise Ratio ◽  
Conformation Change ◽  
Membrane Protein Complex ◽  
H Nmr ◽  
Highly Efficient ◽  
Membrane Protein Complexes

AbstractWhile one dimensional 1H nuclear magnetic resonance (1D 1H-NMR) spectroscopy is one of the most important and convenient method for measuring conformation change in biomacromolecules, characterization of protein dynamics in large membrane protein complexes by 1D 1H-NMR remains challenging, due to the difficulty of spectra assignment, low signal-to-noise ratio (S/N) and the need for large amount of protein. Here we report the site-specific incorporation of 4-trimethylsilyl phenylalanine (TMSiPhe) into proteins, through genetic code expansion in Escherichia coli cells, and the measurement of multiple conformational states in membrane protein complex by 1D 1H-NMR. The unique up-field 1H-NMR chemical shift of TMSiPhe, highly efficient and specific incorporation of TMSiPhe enabled facile assignment of the TMSiPhe 1H-NMR signal, and characterization of multiple conformational state in a 150 kilodalton (kD) membrane protein complex, using only 5 μM of protein and 20 min spectra accumulation time. This highly efficient and convenient methods should be broadly applicable for the investigation of dynamic conformation change of protein complexes.

Conformational Flexibility in Assembly of Photosynthetic Membrane Protein Complexes in vivo

2004 ◽  
Vol 10 (S02) ◽  
pp. 1496-1497
Author(s):  
P A Bullough
Keyword(s):  
Membrane Protein ◽  
Protein Complexes ◽  
Conformational Flexibility ◽  
Photosynthetic Membrane ◽  
Membrane Protein Complexes

Extended abstract of a paper presented at Microscopy and Microanalysis 2004 in Savannah, Georgia, USA, August 1–5, 2004.

Three-dimensional crystallization of membrane proteins

1988 ◽  
Vol 21 (4) ◽  
pp. 429-477 ◽  
Author(s):  
W. Kühlbrandt
Keyword(s):  
High Resolution ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Protein Complexes ◽  
Three Dimensional ◽  
Biological Macromolecules ◽  
X Ray ◽  
X Ray Crystallography ◽  
Membrane Protein Complexes ◽  
Essential Prerequisite

As recently as 10 years ago, the prospect of solving the structure of any membrane protein by X-ray crystallography seemed remote. Since then, the threedimensional (3-D) structures of two membrane protein complexes, the bacterial photosynthetic reaction centres of Rhodopseudomonas viridis (Deisenhofer et al. 1984, 1985) and of Rhodobacter sphaeroides (Allen et al. 1986, 1987 a, 6; Chang et al. 1986) have been determined at high resolution. This astonishing progress would not have been possible without the pioneering work of Michel and Garavito who first succeeded in growing 3-D crystals of the membrane proteins bacteriorhodopsin (Michel & Oesterhelt, 1980) and matrix porin (Garavito & Rosenbusch, 1980). X-ray crystallography is still the only routine method for determining the 3-D structures of biological macromolecules at high resolution and well-ordered 3-D crystals of sufficient size are the essential prerequisite.

scholarly journals Results Regarding the Resolving of Membrane-Protein Complexes using Neutron Reflection in Molecular Dynamics

2018 ◽  
Vol 114 (3) ◽  
pp. 341a
Author(s):  
Bradley W. Treece ◽  
Arvind Ramanathan ◽  
Frank Heinrich ◽  
Mathias Lösche
Keyword(s):  
Molecular Dynamics ◽  
Membrane Protein ◽  
Protein Complexes ◽  
Neutron Reflection ◽  
Membrane Protein Complexes
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