Tethering assays for COPI vesicles mediated by golgins.
A method is described that allows the attachment of COPI vesicles and Golgi membranes to glass slides that can then be analyzed using electron microscopy (EM) and immuno-EM methods. Subpopulations of COPI vesicles can be bound selectively using recombinant golgins. Alternatively, COPI vesicles can be attached to prebound Golgi membranes. Marking these vesicles selectively with biotin allows their site of attachment to be identified.
Satoh
Ayano
A
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, USA.
Malsam
Jörg
J
Warren
Graham
G
D000818:Animals; D001324:Autoantigens; D019921:Biotinylation; D022181:COP-Coated Vesicles / Q000502:physiology*; D002458:Cell Fractionation / Q000379:methods; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004268:DNA-Binding Proteins; D005898:Glass; D006056:Golgi Apparatus / Q000502:physiology*; D000077614:Golgi Matrix Proteins; D008565:Membrane Proteins; D051381:Rats; D014157:Transcription Factors
D001324:Autoantigens; C088424:Cux1 protein, rat; D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D000077614:Golgi Matrix Proteins; D008565:Membrane Proteins; D014157:Transcription Factors; C496921:golgin-84, rat