Three-Dimensional Structure of Cholera Toxin Penetrating a Lipid Membrane

Ribi, HO; Ludwig, David S.; Mercer, K. Lynne; Schoolnik, G; Kornberg, R.D.

doi:10.1126/science.3344432

Cited by 165 publications

(77 citation statements)

References 30 publications

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“…Additionally, the inherently random distribution of GM1 is inconsistent with previous work using fluorescent cholera toxin as a "raft marker" in two-color comparisons with fluorescent signaling molecules in live cells (Stauffer and Meyer, 1997). Because cholera toxin is pentameric (Reed et al, 1987;Ribi et al, 1988), it is likely that the fluorescent toxin, like the gold-conjugated toxin, induces the redistribution of GM1 from a dispersed topography to signaling domains that independently accumulate activated receptors. We suggest using fluorescent cholera toxin not as a marker for lipid rafts, but rather as a marker for membrane regions that are specialized for signal propagation and endocytosis.…”

Section: Aggregated Gm1 and Fc⑀ri But Not Thy-1 Concentrate In Eleccontrasting

confidence: 48%

Markers for Detergent-resistant Lipid Rafts Occupy Distinct and Dynamic Domains in Native Membranes

Wilson

Steinberg

Liederman

et al. 2004

MBoC

187

183

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Lipid rafts isolated by detergent extraction and sucrose gradient fractionation from mast cells are enriched for the glycosylphosphatidylinositol-linked protein Thy-1, the ganglioside GM1, palmitoylated LAT, and cross-linked IgE receptors, Fc⑀RI. This study addresses the relationship of fractionation data to the organization of raft markers in native membranes. Immunogold labeling and electron microscopy shows there is little or no colocalization of the raft markers Thy-1, GM1, and LAT with each other or with Fc⑀RI on native membrane sheets prepared from unstimulated cells. External cross-linking of Thy-1 promotes coclustering of Thy-1 with LAT, but not with GM1. Thy-1 and LAT clusters occur on membrane regions without distinctive features. In contrast, external cross-linking of Fc⑀RI and GM1 causes their redistribution to electron-dense membrane patches independently of each other and of Thy-1. The distinctive patches that accumulate cross-linked Fc⑀RI and GM1 also accumulate osmium, a stain for unsaturated lipids, and are sites for coated vesicle budding. Electron microscopy reveals a more complex and dynamic topographical organization of membrane microdomains than is predicted by biochemical analysis of detergent-resistant membranes. INTRODUCTIONOrdered regions of membrane, known as microdomains, lipid rafts, detergent-resistant membranes (DRMs), and other abbreviations are thought to be critical sites of signal propagation and membrane trafficking (Edidin, 1997(Edidin, , 2001Simons and Ikonen, 1997;Anderson, 1998; London, 1998, 2000;Jacobson and Dietrich, 1999;Langlet et al., 2000;Anderson and Jacobson, 2002). Analysis of these regions typically begins with detergent solubilization of whole cells followed by sucrose density gradient centrifugation and the recovery of detergent-resistant membranes from the light fractions of the gradient. The DRMs are enriched for caveolin, glycosylphosphatidylinositol-linked (GPI-linked) proteins, glycosphingolipids, GM1 ganglioside, and cholesterol, suggesting that these components are associated in the liquid-ordered (l o ) phase of the lipid bilayer (Schroeder et al., 1994;Ahmed et al., 1997). The interpretation of gradient centrifugation experiments remains controversial. There is evidence that detergents may force associations between components that are not colocalized in intact cells (Mayor and Maxfield, 1995), and fractionation results are known to be dramatically altered by varying the concentration of Triton X-100 (Field et al., 1999;Parolini et al., 1999), by use of alternative detergents (Montixi et al., 1998;Surviladze et al., 1998) or by omission of detergent altogether (Ilangumaran et al., 1999;Harder and Kuhn, 2000;Surviladze et al., 2001). Methods to observe membrane segregation in situ have also generated controversy. Results based on light and fluorescence microscopy of proteins and lipids, including refinements of the established fluorescence recovery after photobleaching methods and new single particle tracking methods, have led some investigators to co...

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Section: Aggregated Gm1 and Fc⑀ri But Not Thy-1 Concentrate In Eleccontrasting

confidence: 48%

Markers for Detergent-resistant Lipid Rafts Occupy Distinct and Dynamic Domains in Native Membranes

Wilson

Steinberg

Liederman

et al. 2004

MBoC

187

183

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“…The clustering of some lipids, such as the binding of cholera toxin to GM1 (Ribi et al 1988) or of Shiga toxin to Gb3 (Ling et al 1998), may have an entropic effect on the sorting of other lipids in the membrane, such as that of sphingolipids discussed earlier. Generally, clustering a certain type of lipid by a protein or even actin filaments can induce phase separation in a homogeneous membrane even if this component is present in a very small quantity (Hammond et al 2005;Liu and Fletcher 2006;Römer et al 2010) and (Safouane et al 2010).…”

Section: Protein Binding Enhances Lipid Sortingmentioning

confidence: 99%

Curvature-Driven Lipid Sorting in Biomembranes

Callan-Jones¹,

Sorre²,

Bassereau³

2011

Cold Spring Harbor Perspectives in Biology

129

128

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It has often been suggested that the high curvature of transport intermediates in cells may be a sufficient means to segregate different lipid populations based on the relative energy costs of forming bent membranes. In this review, we present in vitro experiments that highlight the essential physics of lipid sorting at thermal equilibrium: It is driven by a trade-off between bending energy, mixing entropy, and interactions between species. We collect evidence that lipid sorting depends strongly on lipid -lipid and protein-lipid interactions, and hence on the underlying composition of the membrane and on the presence of bound proteins.

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“…This technique has already proven to be very powerful for structural studies of proteins interacting with membranes, enabling the resolution of their structure in the membrane-bound state [10][11][12][13]. In addition, this approach has the potential for high-resolution structure determination by electron crystallography [14] provided that well-ordered 2D crystals are obtained [15].…”

Section: Introductionmentioning

confidence: 99%

Structure of membrane‐bound human factor Va

Stoylova¹,

Mann²,

Brisson³

1994

FEBS Letters

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Coagulation factor Va is an essential cofactor which combines with the serine protease factor Xa on a phospholipid surface to form the prothrombinase complex. In the present study, the structure of factor Va interacting with lipid surfaces containing phosphatidylserine was studied by electron microscopy. Two-dimensional crystals of factor Va were obtained on planar lipid films under quasi-physiological conditions. The two-dimensional projected structure of factor Va was calculated at a resolution of 2 nm, revealing dimers of factor Va arranged on the surface lattice with the symmetry of the plane group p2. Average unit cell dimensions are a = 14.4 nm, b = 8.8 nm, ~, = 107 °. Each factor Va molecule presents two distinct domains of protein density consisting of one small domain, of 3 nm in diameter, connected to a larger domain of about 6 nm x 4.5 nm. The projected structure of factor Va covers an area equivalent to about fifty phospholipid molecules. In addition, edge-on views of factor Va molecules bound to liposomes reveal a globular structure connected through a thin stem to the liposome surface. A three-dimensional model of membrane-bound factor Va is proposed.

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Three-Dimensional Structure of Cholera Toxin Penetrating a Lipid Membrane

Cited by 165 publications

References 30 publications

Markers for Detergent-resistant Lipid Rafts Occupy Distinct and Dynamic Domains in Native Membranes

Markers for Detergent-resistant Lipid Rafts Occupy Distinct and Dynamic Domains in Native Membranes

Curvature-Driven Lipid Sorting in Biomembranes

Structure of membrane‐bound human factor Va

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