Real-time assays for the assembly-disassembly cycle of COP coats on liposomes of defined size.
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.
Bigay
Joëlle
J
Institut de Pharmacologie, Moleculaire et Cellulaire, CNRS, Valbonne, France.
Antonny
Bruno
B
D020823:ADP-Ribosylation Factor 1 / Q000502:physiology*; D022181:COP-Coated Vesicles / Q000502:physiology*; D002499:Centrifugation, Density Gradient / Q000379:methods; D005453:Fluorescence; D020662:Guanine Nucleotide Exchange Factors / Q000502:physiology; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008027:Light; D008081:Liposomes / Q000378:metabolism*; D020559:Monomeric GTP-Binding Proteins / Q000502:physiology*; D011487:Protein Conformation; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D012542:Scattering, Radiation; D033921:Vesicular Transport Proteins
D020662:Guanine Nucleotide Exchange Factors; D008081:Liposomes; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; D020823:ADP-Ribosylation Factor 1; D020559:Monomeric GTP-Binding Proteins; C077395:SAR1 protein, S cerevisiae