Functional analysis of Arl1 and golgin-97 in endosome-to-TGN transport using recombinant Shiga toxin B fragment

Methods Enzymol. 2005:404:442-53. doi: 10.1016/S0076-6879(05)04039-5.

Abstract

A direct transport route from early/recycling endosome (EE/RE) to the trans-Golgi network (TGN) is exploited by Shiga toxin (Mallard et al., 1998) and TGN38 (Ghosh et al., 1998). To facilitate the study of this pathway, both in vivo and in vitro transport assays using recombinant Shiga toxin B fragments (STxB) as protein cargos have facilitated the analysis of this transport event (Johannes et al., 1997; Mallard et al., 1998, 2002; Tai et al., 2004). We describe here the application of these assays to study the role of a small GTPase Arl1 and its effector golgin-97 in this transport process.

MeSH terms

  • ADP-Ribosylation Factors / physiology*
  • Animals
  • Autoantigens / physiology*
  • Endosomes / physiology*
  • Golgi Apparatus / physiology*
  • Golgi Matrix Proteins
  • HeLa Cells
  • Humans
  • Membrane Proteins / physiology*
  • Protein Structure, Tertiary
  • Protein Subunits / metabolism*
  • Protein Transport / drug effects
  • RNA, Small Interfering
  • Rats
  • Recombinant Fusion Proteins
  • Shiga Toxin 2 / metabolism*

Substances

  • Autoantigens
  • Golgi Matrix Proteins
  • Golgi complex autoantigen, 97-kDa
  • Membrane Proteins
  • Protein Subunits
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Shiga Toxin 2
  • shiga toxin 2 B subunit
  • ADP-ribosylation factor related proteins
  • ADP-Ribosylation Factors