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Structure of the Sec23/24–Sar1 pre-budding complex of the COPII vesicle coat

Nature volume 419, pages 271–277 (2002)Cite this article

Abstract

COPII-coated vesicles form on the endoplasmic reticulum by the stepwise recruitment of three cytosolic components: Sar1–GTP to initiate coat formation, Sec23/24 heterodimer to select SNARE and cargo molecules, and Sec13/31 to induce coat polymerization and membrane deformation. Crystallographic analysis of the Saccharomyces cerevisiae Sec23/24–Sar1 complex reveals a bow-tie-shaped structure, 15 nm long, with a membrane-proximal surface that is concave and positively charged to conform to the size and acidic-phospholipid composition of the COPII vesicle. Sec23 and Sar1 form a continuous surface stabilized by a non-hydrolysable GTP analogue, and Sar1 has rearranged from the GDP conformation to expose amino-terminal residues that will probably embed in the bilayer. The GTPase-activating protein (GAP) activity of Sec23 involves an arginine side chain inserted into the Sar1 active site. These observations establish the structural basis for GTP-dependent recruitment of a vesicular coat complex, and for uncoating through coat-controlled GTP hydrolysis.

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Figure 1: Structure of the Sec23/24–Sar1 pre-budding complex.
Figure 2: Domain structure and relatedness of Sec23 and Sec24.
Figure 3: Surface features of the Sec23/24 complex a, Molecular surface coloured according to electrostatic potential50: negative potential is red and positive potential blue.
Figure 4: Conformational switching and protein–protein interactions.
Figure 5: GTP-dependent coat recruitment and coat-dependent GTP hydrolysis a, Stereo view showing interactions between Sar1–GppNHp (pink) and Sec23 (coloured as in Fig. 2).

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Acknowledgements

We thank E. Mossessova for the Sar1 expression construct, J. Walker for assistance and advice on data collection and processing, C. Heaton for use of synchrotron facilities at CHESS, and M. Becker at NSLS. This work was supported by grants from the National Institutes of Health, the Howard Hughes Medial Institute, and the Pew Scholars Program in the Biomedical Sciences.

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  1. Howard Hughes Medical Institute and the Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York, 10021, USA

    Xiping Bi, Richard A. Corpina & Jonathan Goldberg

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Correspondence to Jonathan Goldberg.

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Bi, X., Corpina, R. & Goldberg, J. Structure of the Sec23/24–Sar1 pre-budding complex of the COPII vesicle coat. Nature 419, 271–277 (2002). https://doi.org/10.1038/nature01040

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