Coat Assembly Directs v-SNARE Concentration into Synthetic COPII Vesicles

COPII proteins are required to create transport vesicles and to select cargo molecules for transit from the ER. A reconstituted liposome budding reaction was used to detect the capture and concentration of membrane-associated v-SNARE molecules into synthetic COPII vesicles. A novel glutathione-phosp...

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Published in	Molecular cell Vol. 2; no. 5; pp. 703 - 708
Main Authors	Matsuoka, Ken, Morimitsu, Yasujiro, Uchida, Koji, Schekman, Randy
Format	Journal Article
Language	English
Published	United States Elsevier Inc 01.11.1998
Subjects	Biological Transport Carrier Proteins - physiology Centrifugation, Density Gradient Chromatography, Thin Layer Coated Vesicles - metabolism COPII protein Cross-Linking Reagents Cytoplasm Fungal Proteins - metabolism Glutathione - metabolism Glutathione Transferase Guanine Nucleotide Exchange Factors Liposomes - chemical synthesis Liposomes - metabolism Membrane Glycoproteins - metabolism Membrane Proteins - metabolism Phosphatidylethanolamines - metabolism Phosphoproteins - physiology Qa-SNARE Proteins Qb-SNARE Proteins R-SNARE Proteins Receptors, Cell Surface - metabolism Recombinant Fusion Proteins - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins SNARE Proteins Spectrometry, Mass, Fast Atom Bombardment v-SNARE protein Vesicular Transport Proteins
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ISSN	1097-2765 1097-4164 1097-4164
DOI	10.1016/S1097-2765(00)80168-9

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Summary:	COPII proteins are required to create transport vesicles and to select cargo molecules for transit from the ER. A reconstituted liposome budding reaction was used to detect the capture and concentration of membrane-associated v-SNARE molecules into synthetic COPII vesicles. A novel glutathione-phosphatidylethanolamine conjugate (Glut-PE) was synthesized and incorporated into chemically defined liposomes to provide binding sites for GST hybrid proteins. Large liposomes containing bound cytoplasmic domains of the v-SNAREs, Sec22p or Bos1p, or of the ER resident proteins, Sec12p and Ufe1p, were exposed to COPII proteins and GMP-PNP. v-SNAREs but not resident proteins were concentrated in synthetic COPII vesicles generated from donor liposomes. We conclude that COPII proteins are necessary and sufficient for cargo selection and vesicle morphogenesis.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	1097-2765 1097-4164 1097-4164
DOI:	10.1016/S1097-2765(00)80168-9