TY - JOUR
T1 - COPII-coated vesicle formation reconstituted with purified coat proteins and chemically defined liposomes
AU - Matsuoka, Ken
AU - Orci, Lelio
AU - Amherdt, Mylène
AU - Bednarek, Sebastian Y.
AU - Hamamoto, Susan
AU - Schekman, Randy
AU - Yeung, Thomas
N1 - Funding Information:
We thank Robert Lesch for COPII proteins and Jeremy Thorner and Jonathan A. Ellman for sharing their equipment. We thank Anne Spang and Sebastian Springer for improving the manuscript and members of Schekman lab for discussion and encouragement. This work was supported by the HHMI (R. S.), the Swiss National Fund, and the HFSP (L. O.). K. M. is a visiting scholar from the School of Agricultural Sciences, Nagoya University.
PY - 1998/4/17
Y1 - 1998/4/17
N2 - COPII vesicle formation requires only three coat assembly subunits: Sarl p, Sac13/31 p, and Sec23/24p. PI 4-phosphate or PI 4,5-bisphosphate is required for the binding of these proteins to liposomes. The GTP-bound form of Sar1p recruits Sec23/24p to the liposomes as well as to the ER membranes, and this Sar1p-Sec23/24p complex is required for the binding of Sec13/31p. Ultrastructural analysis shows that the binding of COPII coat proteins to liposomes results in coated patches, coated buds, and coated vesicles of 50- 9 nm in diameter. Budding proceeds without rupture of the donor liposome or vesicle product. These observations suggest that the assembly of the COPII coat on the ER occurs by a sequential binding of coat proteins to specific lipids and that this assembly promotes the budding of COPII-coated vesicles.
AB - COPII vesicle formation requires only three coat assembly subunits: Sarl p, Sac13/31 p, and Sec23/24p. PI 4-phosphate or PI 4,5-bisphosphate is required for the binding of these proteins to liposomes. The GTP-bound form of Sar1p recruits Sec23/24p to the liposomes as well as to the ER membranes, and this Sar1p-Sec23/24p complex is required for the binding of Sec13/31p. Ultrastructural analysis shows that the binding of COPII coat proteins to liposomes results in coated patches, coated buds, and coated vesicles of 50- 9 nm in diameter. Budding proceeds without rupture of the donor liposome or vesicle product. These observations suggest that the assembly of the COPII coat on the ER occurs by a sequential binding of coat proteins to specific lipids and that this assembly promotes the budding of COPII-coated vesicles.
UR - http://www.scopus.com/inward/record.url?scp=18344405156&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=18344405156&partnerID=8YFLogxK
U2 - 10.1016/S0092-8674(00)81577-9
DO - 10.1016/S0092-8674(00)81577-9
M3 - Article
C2 - 9568718
AN - SCOPUS:18344405156
SN - 0092-8674
VL - 93
SP - 263
EP - 275
JO - Cell
JF - Cell
IS - 2
ER -