GPI-Anchored Proteins do not Reside in Ordered Domains in the Live Cell Plasma Membrane

Imaging approaches for analysis of cholesterol distribution and dynamics in the plasma membrane

“…FCS experiments have shown that lipids move with a slower diffusion constant in ordered domains in GPMVs [210] and in the PM [211] than in disordered domains. KK114-PEG-Chol which has been shown to partition equally between the lo and ld phases in GUVs [212] and GPMVs [207,213] [214,215]. Thus, the reduced mobility of CD59 may be explained by the presence of the immobilized mGFP-GPIs.…”

“…For example, mGFP-GPI, a GPI anchored monomeric GFP protein, is a classical marker, known to partition into the lo-like phase in GPMVs [207]. The Schütz group performed an elegant experiment combining micropatterning and SPT to determine whether microdomains packed with mGFP-GPI caused formation of ordered raft-like domains [207]. Briefly, cells expressing mGFP-GPI were grown on micropatterns of mGFP-GPI antibody to form micron sized mGFP-GPI domains in the PM.…”

Protein–lipid interactions critical to replication of the influenza A virus

“…However, the behavior of GPI in cell membranes was recently revisited using micropatterning and single molecule tracking in living cells. These studies did not find that GPI-anchored proteins were targeted to artificially formed GPIanchored GFP domains [21]. Other experiments have directly shown that HA does not reside in L o domains: HA reconstituted in GUVs or HA present in giant plasma membrane vesicles (GPMVs) was found in the L d domain or in both L o and L d [22,23].…”

Adherens Junctions Revisualized: Organizing Cadherins as Nanoassemblies

“…Studies in model membranes demonstrated that interactions between sphingolipids and cholesterol could form tightly packed regions with a specific liquid crystalline organization (the liquid-ordered, L O , phase) (Veatch and Keller, 2002). Since this lipid raft concept was first proposed (Simons and Ikonen, 1997), a consensus has now emerged that-rather than forming large stable platforms of cholesterol, sphingolipids, and associated proteins-rafts constitute highly dynamic assemblies of a few molecules, i.e., small short-lived (<1 s) nanoassemblies (Eggeling et al, 2009;Kusumi et al, 2012;Rao and Mayor, 2014;Simons and Gerl, 2010;Zhou and Hancock, 2015), though it is important to note that the existence of ordered membrane phases in living cells is still a matter of debate (Sevcsik et al, 2015).…”