Neurochemical Research

, Volume 37, Issue 6, pp 1325–1334

Spatial Organization and Stoichiometry of N-Terminal Domain-Mediated Glycosyltransferase Complexes in Golgi Membranes Determined by Fret Microscopy

  • Mariana L. Ferrari
  • Guillermo A. Gomez
  • Hugo J. F. Maccioni
Original Paper

DOI: 10.1007/s11064-012-0741-1

Cite this article as:
Ferrari, M.L., Gomez, G.A. & Maccioni, H.J.F. Neurochem Res (2012) 37: 1325. doi:10.1007/s11064-012-0741-1

Abstract

The functional link between glycolipid glycosyltransferases (GT) relies on the ability of these proteins to form organized molecular complexes. The organization, stoichiometry and composition of these complexes may impact their sorting properties, sub-Golgi localization, and may determine relative efficiency of GT in different glycolipid biosynthetic pathways. In this work, by using Förster resonance energy transfer microscopy in live CHO-K1 cells, we investigated homo- and hetero-complex formation by different GT as well as their spatial organization and molecular stoichiometry on Golgi membranes. We find that GalNAcT and GalT2 Ntd are able to form hetero-complexes in a 1:2 molar ratio at the trans-Golgi network and that GalT2 but not GalNAcT forms homo-complexes. Also, GalNAcT/GalT2 complexes exhibit a stable behavior reflected by its clustered lateral organization. These results reveals that particular topological organization of GTs may have functional implications in determining the composition of glycolipids in cellular membranes.

Keywords

Golgi complexGlycosyltransferase complexesLive cell FRET microscopyGlycolipids

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Mariana L. Ferrari
    • 1
  • Guillermo A. Gomez
    • 1
    • 2
  • Hugo J. F. Maccioni
    • 1
  1. 1.Departamento de Química Biológica, Facultad de Ciencias Químicas, Centro de Investigaciones en Química Biológica de Córdoba, CIQUIBIC (UNC-CONICET)Universidad Nacional de CórdobaCórdobaArgentina
  2. 2.Division of Molecular Cell Biology, Institute for Molecular BioscienceThe University of QueenslandSt. Lucia, BrisbaneAustralia