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Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.
TLDR
A discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) system for the separation of proteins in the range from 1 to 100 kDa is described, and the omission of glycine and urea prevents disturbances which might occur in the course of subsequent amino acid sequencing. Expand
Supercomplexes in the respiratory chains of yeast and mammalian mitochondria
TLDR
A fraction of total bovine ATP synthase was isolated in dimeric form, suggesting that a dimeric state is not limited to S.cerevisiae, but also exists in mammalian mitochondria, which guided us to present a model for a network of respiratory chain complexes: a ‘respirasome’. Expand
Tricine–SDS-PAGE
TLDR
A protocol for Tricine–SDS-PAGE is described, which includes efficient methods for Coomassie blue or silver staining and electroblotting, thereby increasing the versatility of the approach. Expand
Analysis of molecular masses and oligomeric states of protein complexes by blue native electrophoresis and isolation of membrane protein complexes by two-dimensional native electrophoresis.
TLDR
A two-dimensional native technique was developed that is suitable for preparation of highly pure membrane protein complexes and was applied to analysis of cytochrome bc/bf complexes. Expand
Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form.
TLDR
The percentage recovery of functional activity depended on the respective protein complex studied and was zero for some complexes, but almost quantitative for others, and the recovery of all respiratory chain complexes was almost quantitative. Expand
Blue native PAGE
TLDR
This protocol describes methodology to perform BN-PAGE followed by (i) native extraction or native electroblotting of separated proteins, or (ii) a second dimension of tricine-SDS-PA GE or modified BN, followed by a third dimension of isoelectric focusing (IEF) for the separation of subunits of complexes. Expand
Yeast mitochondrial F1F0‐ATP synthase exists as a dimer: identification of three dimer‐specific subunits
TLDR
It is speculated that the F1F0‐ATP synthase of yeast, like the other complexes of oxidative phosphorylation, form supracomplexes to optimize transduction of energy and to enhance the stability of the complex in the membrane. Expand
High Resolution Clear Native Electrophoresis for In-gel Functional Assays and Fluorescence Studies of Membrane Protein Complexes*
TLDR
A substituted Coomassie dye was substituted in the cathode buffer of blue native electrophoresis by non-colored mixtures of anionic and neutral detergents that imposed a charge shift on the membrane proteins to enhance their anodic migration and improved membrane protein solubility during electrophoreis. Expand
Cardiolipin Stabilizes Respiratory Chain Supercomplexes*
TLDR
It is shown that a cardiolipin-deficient strain harbored almost inactive resting cytochrome c oxidase in the membrane and Transition to the fully active pulsed state occurred on a minute time scale. Expand
AIF deficiency compromises oxidative phosphorylation
TLDR
It is shown that human or mouse cells lacking AIF as a result of homologous recombination or small interfering RNA exhibit high lactate production and enhanced dependency on glycolytic ATP generation, due to severe reduction of respiratory chain complex I activity. Expand
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