Transverse distribution of plasma membrane bilayer cholesterol: Picking sides

Steck, Theodore L.; Lange, Yvonne

doi:10.1111/tra.12586

Cited by 101 publications

(92 citation statements)

References 137 publications

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“…The most abundant glycolipid in human RBCs is globoside (Gb4), estimated to be <5% of all lipids(51); PIP 2 is the most abundant phosphoinositide, estimated at <1% (52). Similarly, while the total concentration of cholesterol is quantified (~40 mol%), its distribution between leaflets is not accessible by our methodology and remains controversial (53). Classical measurements found conflicting results (23,54), and these contradictions are mirrored in more recent work, with some groups reporting inner leaflet enrichment of cholesterol (55,56) and others the opposite (57).…”

Section: Rbcsmentioning

confidence: 99%

The mammalian plasma membrane is defined by transmembrane asymmetries in lipid unsaturation, leaflet packing, and protein shape

Lorent

Levental

Ganesan

et al. 2019

Preprint

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A fundamental feature of cellular plasma membranes (PM) is the asymmetric distribution of lipids between the bilayer leaflets. This asymmetry is central to cellular physiology, regulating signaling, apoptosis, coagulation, and cell-cell fusion. While the broad transbilayer distributions of some lipid types are well established, the detailed lipidomic asymmetry of the PM has not been characterized and thus the compositions of the individual leaflets remain poorly understood. Further, how these compositional differences contribute to structural asymmetries between PM leaflets in living cells is not defined. Here, we report the distinct lipidomes and biophysical properties of both monolayers in living mammalian PMs. Using mass spectrometry coupled to enzymatic digestion, we report the detailed compositions of PM leaflets of human erythrocytes. We find a dramatic asymmetry in phospholipid unsaturation, with the cytoplasmic leaflet being ~2-fold more unsaturated than the exoplasmic. Atomistic simulations of lipid mixtures compiled from the lipidomic observations suggested significant asymmetries in lipid order, packing, and dynamics between the two PM leaflets. These were probed directly in the PM of living cells by Fluorescence Lifetime Imaging Microscopy (FLIM) of leaflet-selective, environment-sensitive probes. The outer PM leaflet is highly packed and ordered, resembling a liquid ordered phase, whereas the inner leaflet is significantly more disordered. This biophysical asymmetry is maintained in the endocytic system. These observations reveal the detailed compositional and biophysical asymmetry of mammalian plasma membranes, elucidating fundamental design principles of living membranes. SIGNIFICANCE STATEMENT:The asymmetric distribution of lipids between the two bilayer leaflets is a fundamental feature of mammalian plasma membranes. Here, we quantify the comprehensive lipidomes of the two PM leaflets in living mammalian cells and examine their consequences on leaflet physical properties. Using computer simulations, we predict distinct biophysical features for the two leaflets and confirm these with a variety of spectroscopic and microscopic techniques. We find that the outer leaflet of the membrane contains many more fully saturated lipids, which endows it with higher lipid packing and lower diffusivity.

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Section: Rbcsmentioning

confidence: 99%

The mammalian plasma membrane is defined by transmembrane asymmetries in lipid unsaturation, leaflet packing, and protein shape

Lorent

Levental

Ganesan

et al. 2019

Preprint

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“…Heijnen et al demonstrated an enrichment of Chol in filopodia of adhering platelets using a nonhemolytic derivative (BCθ) of Chol‐specific bacterial toxin, perfringolysin O (PFO, θ‐toxin) . In the plasma membrane of mammalian cells, PFO binds when Chol concentration exceeds 35% .…”

Section: Introductionmentioning

confidence: 99%

Cholesterol asymmetry at the tip of filopodia during cell adhesion

et al. 2020

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During adhesion, cells develop filopodia to facilitate the attachment to the extracellular matrix. The small guanosine triphosphate (GTP)-binding protein, Cdc42, plays a central role in the formation of filopodia. It has been reported that Cdc42 activity is regulated by cholesterol (Chol). We examined Chol distribution in filopodia using Chol-binding domain 4 (D4) fragment of bacterial toxin, perfringolysin O that senses high membrane concentration of Chol. Our results indicate that fluorescent D4 was enriched at the tip of the outer leaflet of filopodia in the initiation phase of cell adhesion. This enrichment was accompanied by a defect of D4 labeling in the inner leaflet. Steady phase adhered cell experiment indicated that both Cdc42 and ATP-binding cassette transporter, ABCA1, were involved in the binding of D4 to the cell surface. Depletion of Chol activated Cdc42. Our results suggest that asymmetric distribution of Chol at the tip of filopodia induces activation of Cdc42, and thus, facilitates filopodia formation. K E Y W O R D S ABCA1, Cdc42, cholesterol probe, lipid asymmetry 6186 | KISHIMOTO eT al.

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“…Cholesterol is unevenly distributed within mammalian cells, and the cholesterol content in different membrane compartments varies markedly. Further, cholesterol is also distributed unevenly in the bilayer membrane, both laterally and between the inner and outer leaflets of the bilayer (1)(2)(3).…”

Section: Introductionmentioning

confidence: 99%

The Affinity of Sterols for Different Phospholipid Classes and Its Impact on Lateral Segregation

Nyholm

Jaikishan

Engberg

et al. 2019

Biophysical Journal

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Cholesterol is an essential molecule in the membranes of mammalian cells. It is known to be distributed heterogeneously within the cells, between the bilayer leaflets, as well as between lateral domains within the bilayer. However, we do not know exactly how cholesterol is distributed and what forces drive this sorting process because it extremely difficult to study using currently available methods. To further elucidate this distribution, we measured how cholesterol partitions between different phospholipid (PL) environments using different methods based on cholesterol, TopFluor-cholesterol, and cholesta-5,7,9(11)-triene-3-b-ol. Based on the obtained relative partition coefficients, we made predictions regarding how cholesterol would be distributed between lateral domains and between the inner and outer leaflets of the plasma membrane. In addition, using a trans-parinaric acid fluorescence-based method, we tested how cholesterol could influence lateral segregation through its interaction with unsaturated PLs with different headgroups. The results showed that the lower the affinity of cholesterol was for the different unsaturated PLs, the more cholesterol stimulated lateral segregation in a ternary bilayer of unsaturated PL/ N-palmitoyl-D-erythro-sphingomyelin and cholesterol. Overall, the results indicate that both the distribution of cholesterol between different lipid environments and the impact of cholesterol on lateral segregation can be predicted relatively accurately from determined relative partition coefficients.

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Transverse distribution of plasma membrane bilayer cholesterol: Picking sides

Cited by 101 publications

References 137 publications

The mammalian plasma membrane is defined by transmembrane asymmetries in lipid unsaturation, leaflet packing, and protein shape

The mammalian plasma membrane is defined by transmembrane asymmetries in lipid unsaturation, leaflet packing, and protein shape

Cholesterol asymmetry at the tip of filopodia during cell adhesion

The Affinity of Sterols for Different Phospholipid Classes and Its Impact on Lateral Segregation

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