Proteomic characterization of golgi membranes enriched from Arabidopsis suspension cell cultures

Sara Fasmer Hansen, Berit Ebert, Carsten Rautengarten, Joshua L. Heazlewood

1 Citation (Scopus)

Abstract

The plant Golgi apparatus has a central role in the secretory pathway and is the principal site within the cell for the assembly and processing of macromolecules. The stacked membrane structure of the Golgi apparatus along with its interactions with the cytoskeleton and endoplasmic reticulum has historically made the isolation and purification of this organelle difficult. Density centrifugation has typically been used to enrich Golgi membranes from plant microsomal preparations, and aside from minor adaptations, the approach is still widely employed. Here we outline the enrichment of Golgi membranes from an Arabidopsis cell suspension culture that can be used to investigate the proteome of this organelle. We also provide a useful workflow for the examination of proteomic data as the result of multiple analyses. Finally, we highlight a simple technique to validate the subcellular localization of proteins by fluorescent tags after their identification by tandem mass spectrometry.

Original languageEnglish
Title of host publicationThe golgi complex : methods and protocols
EditorsWilliam J. Brown
Number of pages19
PublisherSpringer
Publication date1 Sept 2016
Pages91-109
Chapter8
ISBN (Print)978-1-4939-6461-1
ISBN (Electronic)978-1-4939-6463-5
DOIs
Publication statusPublished - 1 Sept 2016
SeriesMethods in Molecular Biology
Volume1496
ISSN1064-3745

Keywords

  • Journal Article

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