Biochemical sub-fractionation of the mammalian Golgi apparatus

Tomohiko Taguchi, Marc Pypaert, Graham Warren

Research output: Contribution to journalReview articlepeer-review

17 Citations (Scopus)

Abstract

We have exploited the breakdown of the Golgi apparatus that occurs during mitosis to isolate subfractions using immuno-affinity methods. Rat liver Golgi stacks were treated with mitotic cytosol from HeLa cells, and the fragments were then incubated with antibodies immobilized on magnetic beads. Antibodies against the cis-Golgi marker, GM130, bound membranes that were depleted in the trans-Golgi network marker, TGN38, whereas antibodies against the cytoplasmic tail of TGN38 did the reverse. A range of other Golgi enzymes, SNAREs and tethers were also tested and were found to bind to anti-GM130 antibodies to an extent that reflected their proximity to cis-cisternae as determined by other techniques. This method should provide a useful complement to the immuno-EM methods presently used to map the Golgi apparatus.

Original languageEnglish
Pages (from-to)344-352
Number of pages9
JournalTraffic
Volume4
Issue number5
DOIs
Publication statusPublished - 2003 May 1
Externally publishedYes

Keywords

  • Fractionation
  • Golgi
  • Golgin
  • Mitosis
  • SNARE
  • Tether

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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