Abstract
The assembly-disassembly cycle of COPI and COPII coats is controlled by the GTPase cycle of the small G proteins Arf1 and Sar. We describe here two spectroscopic assays that enable real-time studies of some elementary steps of coat assembly and disassembly on artificial liposomes of defined composition and curvature. A flotation assay to assess the effect of membrane curvature on protein adsorption to liposomes is also presented.
MeSH terms
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ADP-Ribosylation Factor 1 / physiology*
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COP-Coated Vesicles / physiology*
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Centrifugation, Density Gradient / methods
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Fluorescence
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Guanine Nucleotide Exchange Factors / physiology
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Guanosine Diphosphate / metabolism
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Guanosine Triphosphate / metabolism
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Light
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Liposomes / metabolism*
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Monomeric GTP-Binding Proteins / physiology*
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Protein Conformation
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Saccharomyces cerevisiae Proteins / physiology*
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Scattering, Radiation
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Vesicular Transport Proteins
Substances
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Guanine Nucleotide Exchange Factors
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Liposomes
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Saccharomyces cerevisiae Proteins
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Vesicular Transport Proteins
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Guanosine Diphosphate
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Guanosine Triphosphate
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ADP-Ribosylation Factor 1
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Monomeric GTP-Binding Proteins
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SAR1 protein, S cerevisiae