Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016:1496:63-74.
doi: 10.1007/978-1-4939-6463-5_6.

Reconstitution of COPI Vesicle and Tubule Formation

Seung-Yeol Park  1   2 , Jia-Shu Yang  1   2 , Victor W Hsu  3   4   5
Affiliations

Reconstitution of COPI Vesicle and Tubule Formation

Seung-Yeol Park et al. Methods Mol Biol. 2016.

Abstract

The Golgi complex plays a central role in the intracellular sorting of proteins. Transport through the Golgi in the anterograde direction has been explained by cisternal maturation, while transport in the retrograde direction is attributed to vesicles formed by the coat protein I (COPI) complex. A more detailed understanding of how COPI acts in Golgi transport is being achieved in recent years, due in large part to a COPI reconstitution system. Through this approach, the mechanistic complexities of COPI vesicle formation are being elucidated. This approach has also uncovered a new mode of anterograde transport through the Golgi, which involves COPI tubules connecting the Golgi cisternae. We describe in this chapter the reconstitution of COPI vesicle and tubule formation from Golgi membrane.

Keywords: ARF; COPI; Golgi complex; Tubule formation; Vesicle formation.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Schematic of the two-stage incubation system to reconstitute COPI vesicles and tubules. In the first stage, Golgi membrane is incubated with purified ARF1 and coatomer. In the second stage, the incubated Golgi membrane is further incubated with purified ARFGAP1 and BARS to form COPI vesicles. For COPI tubule formation, the second-stage incubation involves the further addition of either purified cPLA2α or Cdc42.
See this image and copyright information in PMC

References

    1. Glick BS, Nakano A (2009) Membrane Traffic Within the Golgi Apparatus. Annual Review of Cell and Developmental Biology 25 (1):113–132 - PMC - PubMed
    1. Nakano A, Luini A (2010) Passage through the Golgi. Current Opinion in Cell Biology 22 (4):471–478 - PubMed
    1. Malhotra V, Serafini T, Orci L, Shepherd JC, Rothman JE (1989) Purification of a novel class of coated vesicles mediating biosynthetic protein transport through the Golgi stack. Cell 58 (2):329–336 - PubMed
    1. Waters MG, Serafini T, Rothman JE (1991) ‘Coatomer’: a cytosolic protein complex containing subunits of non-clathrin-coated Golgi transport vesicles. Nature 349 (6306):248–251 - PubMed
    1. Donaldson JG, Cassel D, Kahn RA, Klausner RD (1992) ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein beta-COP to Golgi membranes. Proc Natl Acad Sci USA 89 (14):6408–6412 - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources