Protein flexibility is required for vesicle tethering at the Golgi

  1. Pak-yan Patricia Cheung
  2. Charles Limouse
  3. Hideo Mabuchi
  4. Suzanne R Pfeffer  Is a corresponding author
  1. Stanford University School of Medicine, United States
  2. Stanford University, United States

Abstract

The Golgi is decorated with coiled-coil proteins that may extend long distances to help vesicles find their targets. GCC185 is a trans Golgi-associated protein that captures vesicles inbound from late endosomes. Although predicted to be relatively rigid and highly extended, we show that flexibility in a central region is required for GCC185's ability to function in a vesicle tethering cycle. Proximity ligation experiments show that that GCC185's N-and C-termini are within <40nm of each other on the Golgi. In physiological buffers without fixatives, atomic force microscopy reveals that GCC185 is shorter than predicted, and its flexibility is due to a central bubble that represents local unwinding of specific sequences. Moreover, 85% of the N-termini are splayed, and the splayed N-terminus can capture transport vesicles in vitro. These unexpected features support a model in which GCC185 collapses onto the Golgi surface, perhaps by binding to Rab GTPases, to mediate vesicle tethering.

Article and author information

Author details

  1. Pak-yan Patricia Cheung

    Department of Biochemistry, Stanford University School of Medicine, Stanford, United States
    Competing interests
    No competing interests declared.
  2. Charles Limouse

    Department of Applied Physics, Stanford University, Stanford, United States
    Competing interests
    No competing interests declared.
  3. Hideo Mabuchi

    Department of Applied Physics, Stanford University, Stanford, United States
    Competing interests
    No competing interests declared.
  4. Suzanne R Pfeffer

    Department of Biochemistry, Stanford University School of Medicine, Stanford, United States
    For correspondence
    pfeffer@stanford.edu
    Competing interests
    Suzanne R Pfeffer, Reviewing editor, eLife.

Copyright

© 2015, Cheung et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,605
    views
  • 588
    downloads
  • 63
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Citations by DOI

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Pak-yan Patricia Cheung
  2. Charles Limouse
  3. Hideo Mabuchi
  4. Suzanne R Pfeffer
(2015)
Protein flexibility is required for vesicle tethering at the Golgi
eLife 4:e12790.
https://doi.org/10.7554/eLife.12790

Share this article

https://doi.org/10.7554/eLife.12790