Determination of the Membrane Environment of CD59 in Living Cells.

Biomolecules
Gergő FülöpEva Sevcsik

Abstract

The organization and dynamics of proteins and lipids in the plasma membrane, and their role in membrane functionality, have been subject of a long-lasting debate. Specifically, it is unclear to what extent membrane proteins are affected by their immediate lipid environment and vice versa. Studies on model membranes and plasma membrane vesicles indicated preferences of proteins for lipid phases characterized by different acyl chain order; however, whether such phases do indeed exist in live cells is still not known. Here, we refine a previously developed micropatterning approach combined with single molecule tracking to quantify the influence of the glycosylphosphatidylinositol-anchored (GPI-anchored) protein CD59 on its molecular environment directly in the live cell plasma membrane. We find that locally enriched and immobilized CD59 presents obstacles to the diffusion of fluorescently labeled lipids with a different phase-partitioning behavior independent of cell cholesterol levels and type of lipid. Our results give no evidence for either specific binding of the lipids to CD59 or the existence of nanoscopic ordered membrane regions associated with CD59.

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Citations

Jun 28, 2020·FEBS Letters·Maria J SarmentoRadek Šachl
Feb 9, 2019·Frontiers in Chemistry·Marco LindnerEva Sevcsik
Aug 31, 2018·Langmuir : the ACS Journal of Surfaces and Colloids·Roland HagerStefan Howorka

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Methods Mentioned

BETA
fluorescence cross-correlation spectroscopy
glycosylation
Fluorescence
fluorescence microscopy
FCS

Software Mentioned

Matlab
Origin
FlowJo

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