Protein Association with Membrane Rafts

Abstract

Membrane rafts are very small and highly dynamic assemblies in cellular membranes enriched in cholesterol and sphingolipids. Some proteins can functionally associate with rafts: peripheral membrane proteins are incorporated into rafts depending on cues such as the presence of a glycosyl‐phosphatidylinositol (GPI) anchor or S‐acylation (palmitoylation); transmembrane proteins can partition into raft domains depending on specific features within their transmembrane domain. Raft association of membrane proteins was originally defined by their resistance to cold Triton X‐100 extraction, which is however insufficient as the sole criterion – more sophisticated methodology such as fluorescence resonance energy transfer (FRET) has to be employed to determine whether and how a given protein interacts with raft structures.

Key Concepts:

  • Membrane rafts are small, dynamic clusters within biological membranes enriched in cholesterol and sphingolipids.

  • Rafts can be coalesced and stabilised to fulfil a biological function, for example, signal transduction or virus budding.

  • Some proteins are capable of partitioning into raft domains.

  • Raft‐targeting features in proteins are glycosyl‐phosphatidylinositol (GPI) anchors, S‐acylation (palmitoylation) and structural motifs in the transmembrane domain.

  • Assessment of detergent‐resistant membranes (DRM), the original biochemical method to analyse raft association of a protein, is artefact‐prone and therefore not suitable to prove raft involvement in a biological process.

  • More sophisticated methodology such as fluorescence resonance energy transfer (FRET) is needed to decipher raft association of a protein.

Keywords: membrane raft; cholesterol; sphingolipid; glycosyl‐phosphatidylinositol (GPI) anchor; S‐acylation/palmitoylation; detergent‐resistant membranes (DRM); fluorescence microscopy; fluorescence resonance energy transfer (FRET); nanoscopy; model membranes

Figure 1.

Lipids and membrane rafts. (a) The major membrane lipids glycerophospholipids (palmitoyl‐oleoyl‐phosphatidylcholine is shown), sphingolipids and cholesterol with their hydrophilic headgroups indicated by a circle. (b) Sphingolipids and cholesterol have the propensity to form a liquid‐ordered membrane phase.

Figure 2.

Cellular membrane rafts. (a) Rafts in resting cells are very small and dynamic. Some proteins can dynamically partition into these rafts (1, GPI‐anchored protein; 2, double acylated protein; 3, transmembrane protein with a hypothetical lipid binding pocket [blue] and/or palmitoylation), others (4, 5) cannot. (b) Rafts can be coalesced to larger, stabilised platforms to fulfil a biological function. Raft proteins (1, 2, 3) are concentrated in rafts by, for example, ligand binding, effectors (yellow) can bind to these platforms. For some prominent examples for raft proteins (1, 2, 3), see Table .

Figure 3.

Covalent lipid modifications of proteins: myristoylation, S‐acylation and isoprenylation. See Table for details.

close

References

Anderson RG and Jacobson K (2002) A role for lipid shells in targeting proteins to caveolae, rafts, and other lipid domains. Science 296: 1821–1825.

Baumgart T, Hammond AT, Sengupta P et al. (2007) Large‐scale fluid/fluid phase separation of proteins and lipids in giant plasma membrane vesicles. Proceedings of the National Academy of Sciences of the United States of America 104: 3165–3170.

Brown DA and London E (2000) Structure and function of sphingolipid‐ and cholesterol‐rich membrane rafts. Journal of Biological Chemistry 275: 17221–17224.

Brown DA and Rose JK (1992) Sorting of GPI‐anchored proteins to glycolipid‐enriched membrane subdomains during transport to the apical cell surface. Cell 68: 533–544.

Devaux PF and Morris R (2004) Transmembrane asymmetry and lateral domains in biological membranes. Traffic 5: 241–246.

Dietrich C, Bagatolli LA, Volovyk ZN et al. (2001) Lipid rafts reconstituted in model membranes. Biophysical Journal 80: 1417–1428.

Dietzen DJ, Hastings WR and Lublin DM (1995) Caveolin is palmitoylated on multiple cysteine residues. Palmitoylation is not necessary for localization of caveolin to caveolae. Journal of Biological Chemistry 270: 6838–6842.

Edidin M (2003) The state of lipid rafts: from model membranes to cells. Annual Reviews of Biophysics and Biomolecular Structure 32: 257–283.

Eggeling C, Ringemann C, Medda R et al. (2009) Direct observation of the nanoscale dynamics of membrane lipids in a living cell. Nature 457: 1159–1162.

Engel S, Scolari S, Thaa B et al. (2010) FLIM‐FRET and FRAP reveal association of influenza virus haemagglutinin with membrane rafts. Biochemical Journal 425: 567–573.

Epand RM (2006) Cholesterol and the interaction of proteins with membrane domains. Progress in Lipid Research 45: 279–294.

Glebov OO and Nichols BJ (2004) Lipid raft proteins have a random distribution during localized activation of the T‐cell receptor. Nature Cell Biology 6: 238–243.

Goswami D, Gowrishankar K, Bilgrami S et al. (2008) Nanoclusters of GPI‐anchored proteins are formed by cortical actin‐driven activity. Cell 135: 1085–1097.

Hanson MA, Cherezov V, Griffith MT et al. (2008) A specific cholesterol binding site is established by the 2.8 Å structure of the human beta2‐adrenergic receptor. Structure 16: 897–905.

Harder T, Scheiffele P, Verkade P and Simons K (1998) Lipid domain structure of the plasma membrane revealed by patching of membrane components. Journal of Cell Biology 141: 929–942.

Hemler ME (2005) Tetraspanin functions and associated microdomains. Nature Reviews Molecular Cell Biology 6: 801–811.

Hess ST, Gould TJ, Gudheti MV et al. (2007) Dynamic clustered distribution of hemagglutinin resolved at 40 nm in living cell membranes discriminates between raft theories. Proceedings of the National Academy of Sciences of the United States of America 104: 17370–17375.

Hess ST, Kumar M, Verma A et al. (2005) Quantitative electron microscopy and fluorescence spectroscopy of the membrane distribution of influenza hemagglutinin. Journal of Cell Biology 169: 965–976.

Huang B, Babcock H and Zhuang X (2010) Breaking the diffraction barrier: super‐resolution imaging of cells. Cell 143: 1047–1058.

Kaiser HJ, Lingwood D, Levental I et al. (2009) Order of lipid phases in model and plasma membranes. Proceedings of the National Academy of Sciences of the United States of America 106: 16645–16650.

Kellner RR, Baier CJ, Willig KI, Hell SW and Barrantes FJ (2007) Nanoscale organization of nicotinic acetylcholine receptors revealed by stimulated emission depletion microscopy. Neuroscience 144: 135–143.

Kenworthy AK (2008) Have we become overly reliant on lipid rafts? Talking point on the involvement of lipid rafts in T‐cell activation. EMBO Reports 9: 531–535.

Kenworthy AK, Nichols BJ, Remmert CL et al. (2004) Dynamics of putative raft‐associated proteins at the cell surface. Journal of Cell Biology 165: 735–746.

Kordyukova LV, Serebryakova MV, Baratova LA and Veit M (2008) S acylation of the hemagglutinin of influenza viruses: mass spectrometry reveals site‐specific attachment of stearic acid to a transmembrane cysteine. Journal of Virology 82: 9288–9292.

Kordyukova LV, Serebryakova MV, Baratova LA and Veit M (2010) Site‐specific attachment of palmitate or stearate to cytoplasmic versus transmembrane cysteines is a common feature of viral spike proteins. Virology 398: 49–56.

Kusumi A, Shirai YM, Koyama‐Honda I, Suzuki KG and Fujiwara TK (2010) Hierarchical organization of the plasma membrane: investigations by single‐molecule tracking vs. fluorescence correlation spectroscopy. FEBS Letters 584: 1814–1823.

Leser GP and Lamb RA (2005) Influenza virus assembly and budding in raft‐derived microdomains: a quantitative analysis of the surface distribution of HA, NA and M2 proteins. Virology 342: 215–227.

Levental I, Grzybek M and Simons K (2010a) Greasing their way: lipid modifications determine protein association with membrane rafts. Biochemistry 49: 6305–6316.

Levental I, Lingwood D, Grzybek M, Coskun Ü and Simons K (2010b) Palmitoylation regulates raft affinity for the majority of integral raft proteins. Proceedings of the National Academy of Sciences of the United States of America 107: 22050–22054.

Lichtenberg D, Goni FM and Heerklotz H (2005) Detergent‐resistant membranes should not be identified with membrane rafts. Trends in Biochemical Sciences 30: 430–436.

Lingwood D and Simons K (2010) Lipid rafts as a membrane‐organizing principle. Science 327: 46–50.

Melkonian KA, Ostermeyer AG, Chen JZ, Roth MG and Brown DA (1999) Role of lipid modifications in targeting proteins to detergent‐resistant membrane rafts. Many raft proteins are acylated, while few are prenylated. Journal of Biological Chemistry 274: 3910–3917.

Munro S (2003) Lipid rafts: elusive or illusive? Cell 115: 377–388.

Nikolaus J, Scolari S, Bayraktarov E et al. (2010) Hemagglutinin of influenza virus partitions into the nonraft domain of model membranes. Biophysical Journal 99: 489–498.

Owen DM, Williamson D, Rentero C and Gaus K (2009) Quantitative microscopy: protein dynamics and membrane organisation. Traffic 10: 962–971.

Patterson GH, Hirschberg K, Polishchuk RS et al. (2008) Transport through the Golgi apparatus by rapid partitioning within a two‐phase membrane system. Cell 133: 1055–1067.

Pike LJ (2006) Rafts defined: a report on the keystone symposium on lipid rafts and cell function. Journal of Lipid Research 47: 1597–1598.

Prior IA, Harding A, Yan J et al. (2001) GTP‐dependent segregation of H‐ras from lipid rafts is required for biological activity. Nature Cell Biology 3: 368–375.

Prior IA, Muncke C, Parton RG and Hancock JF (2003) Direct visualization of Ras proteins in spatially distinct cell surface microdomains. Journal of Cell Biology 160: 165–170.

Rao M and Mayor S (2005) Use of Förster's resonance energy transfer microscopy to study lipid rafts. Biochimica et Biophysica Acta 1746: 221–233.

Saad JS, Miller J, Tai J et al. (2006) Structural basis for targeting HIV‐1 Gag proteins to the plasma membrane for virus assembly. Proceedings of the National Academy of Sciences of the United States of America 103: 11364–11369.

Scheiffele P, Roth MG and Simons K (1997) Interaction of influenza virus haemagglutinin with sphingolipid‐cholesterol membrane domains via its transmembrane domain. EMBO Journal 16: 5501–5508.

Schuck S and Simons K (2004) Polarized sorting in epithelial cells: raft clustering and the biogenesis of the apical membrane. Journal of Cell Science 117: 5955–5964.

Scolari S, Engel S, Krebs N et al. (2009) Lateral distribution of the transmembrane domain of influenza virus hemagglutinin revealed by time‐resolved fluorescence imaging. Journal of Biological Chemistry 284: 15708–15716.

Sharma P, Varma R, Sarasij RC et al. (2004) Nanoscale organization of multiple GPI‐anchored proteins in living cell membranes. Cell 116: 577–589.

Sharpe HJ, Stevens TJ and Munro S (2010) A comprehensive comparison of transmembrane domains reveals organelle‐specific properties. Cell 142: 158–169.

Shvartsman DE, Kotler M, Tall RD, Roth MG and Henis YI (2003) Differently anchored influenza hemagglutinin mutants display distinct interaction dynamics with mutual rafts. Journal of Cell Biology 163: 879–888.

Simons K and Ikonen E (1997) Functional rafts in cell membranes. Nature 387: 569–572.

Simons K and Vaz WL (2004) Model systems, lipid rafts, and cell membranes. Annual Reviews of Biophysics and Biomolecular Structure 33: 269–295.

Stöckl MT and Herrmann A (2010) Detection of lipid domains in model and cell membranes by fluorescence lifetime imaging microscopy. Biochimica et Biophysica Acta 1798: 1444–1456.

Suomalainen M (2002) Lipid rafts and assembly of enveloped viruses. Traffic 3: 705–709.

Vogel A, Reuther G, Weise K et al. (2009) The lipid modifications of Ras that sense membrane environments and induce local enrichment. Angewandte Chemie International Edition in English 48: 8784–8787.

Zacharias DA, Violin JD, Newton AC and Tsien RY (2002) Partitioning of lipid‐modified monomeric GFPs into membrane microdomains of live cells. Science 296: 913–916.

Further Reading

http://www.nature.com/horizon/livingfrontier/highlights.html

Lingwood D and Simons K (2007) Detergent resistance as a tool in membrane research. Nature Protocols 2: 2159–2165.

van Meer G, Voelker DR and Feigenson GW (2010) Membrane lipids: where they are and how they behave. Nature Reviews Molecular Cell Biology 9: 112–124.

Shevchenko A and Simons K (2010) Lipidomics: coming to grips with lipid diversity. Nature Reviews Molecular Cell Biology 11: 593–598.

Simons K and Gerl MJ (2010) Revitalizing membrane rafts: new tools and insights. Nature Reviews Molecular Cell Biology 11: 688–699.

Tamm LK (ed.) (2005) Protein–Lipid Interactions. From Membrane Domains to Cellular Networks. Weinheim: Wiley‐VCH.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Veit, Michael, and Thaa, Bastian(Jun 2011) Protein Association with Membrane Rafts. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0023404]