Monitoring the diffusion of single heterotrimeric G proteins in supported cell-membrane sheets reveals their partitioning into microdomains

JB Perez, JM Segura, Daniel Abankwa, J Piguet, KL Martinez, H Vogel

Tutkimustuotos: LehtiartikkeliArtikkeliTieteellinenvertaisarvioitu

30 Sitaatiot (Scopus)

Abstrakti

Supported cell-membrane sheets are promising in vitro systems to investigate the properties of membranes with native protein/lipid composition, in particular their sub-compartmentalization and the differential localization of proteins associated to them. While such studies are usually performed using static microscopy techniques, we demonstrate here the potential offered by dynamic diffusion measurements. Whereas the overall fluidity of the lipid bilayer was preserved, the preparation of the membrane sheets led to the selective immobilization of extracellular and transmembrane (TM) glycosylated proteins and the anchored proteins/lipids associated with them. Taking advantage of this, we investigated the association of the G protein Gq with TM proteins, in particular G-protein coupled receptors (GPCRs), by monitoring the changes in diffusion occurring after preparation of the supported membranes. Two fluorescently tagged G alpha q proteins were constructed, which remained either mostly monomeric in the plasma membrane or associated with G beta gamma in heterotrimers. While both constructs diffused similarly in living cells, the preparation of the supported membranes led to the selective immobilization of the heterotrimers with minimal changes of the diffusion of the monomeric Gaq. The diverse mobility of monomeric and heterotrimeric Gaq was a result of their different lipid anchors as demonstrated by monitoring the diffusion of the corresponding anchors alone. We propose that the immobilization of the heterotrimer was caused by its partitioning inside membrane microdomains surrounding GPCRs. (c) 2006 Elsevier Ltd. All rights reserved.
AlkuperäiskieliEi tiedossa
Sivut918–930
Sivumäärä13
JulkaisuJournal of Molecular Biology
Vuosikerta363
Numero5
DOI - pysyväislinkit
TilaJulkaistu - 2006
OKM-julkaisutyyppiA1 Julkaistu artikkeli, soviteltu

Keywords

  • G protein
  • membrane anchors
  • microdomains
  • single-molecule microscopy

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