GPI-anchored proteins are confined in subdiffraction clusters at the apical surface of polarized epithelial cells - Institut Pasteur Access content directly
Journal Articles Biochemical Journal Year : 2017

GPI-anchored proteins are confined in subdiffraction clusters at the apical surface of polarized epithelial cells

Abstract

Spatio-temporal compartmentalization of membrane proteins is critical for the regulation of diverse vital functions in eukaryotic cells. It was previously shown that, at the apical surface of polarized MDCK cells, glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) are organized in small cholesterol-independent clusters of single GPI-AP species (homoclusters), which are required for the formation of larger cholesterol-dependent clusters formed by multiple GPI-AP species (heteroclusters). This clustered organization is crucial for the biological activities of GPI-APs; hence, understanding the spatio-temporal properties of their membrane organization is of fundamental importance. Here, by using direct stochastic optical reconstruction microscopy coupled to pair correlation analysis (pc-STORM), we were able to visualize and measure the size of these clusters. Specifically, we show that they are non-randomly distributed and have an average size of 67 nm. We also demonstrated that polarized MDCK and non-polarized CHO cells have similar cluster distribution and size, but different sensitivity to cholesterol depletion. Finally, we derived a model that allowed a quantitative characterization of the cluster organization of GPI-APs at the apical surface of polarized MDCK cells for the first time. Experimental FRET (fluores-cence resonance energy transfer)/FLIM (fluorescence-lifetime imaging microscopy) data were correlated to the theoretical predictions of the model.
Fichier principal
Vignette du fichier
BCJ-474-4075.pdf (2.8 Mo) Télécharger le fichier
Origin : Publication funded by an institution
Loading...

Dates and versions

pasteur-02078678 , version 1 (25-03-2019)

Licence

Attribution

Identifiers

Cite

Simona Paladino, Stéphanie Lebreton, Mickaël Lelek, Patrizia Riccio, Sergio de Nicola, et al.. GPI-anchored proteins are confined in subdiffraction clusters at the apical surface of polarized epithelial cells. Biochemical Journal, 2017, 474 (24), pp.4075-4090. ⟨10.1042/BCJ20170582⟩. ⟨pasteur-02078678⟩
43 View
89 Download

Altmetric

Share

Gmail Facebook X LinkedIn More