• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biophysjLink to Publisher's site
Biophys J. Nov 2002; 83(5): 2681–2692.
PMCID: PMC1302352

Translational diffusion of individual class II MHC membrane proteins in cells.

Abstract

Single-molecule epifluorescence microscopy was used to observe the translational motion of GPI-linked and native I-E(k) class II MHC membrane proteins in the plasma membrane of CHO cells. The purpose of the study was to look for deviations from Brownian diffusion that might arise from barriers to this motion. Detergent extraction had suggested that these proteins may be confined to lipid microdomains in the plasma membrane. The individual I-E(k) proteins were visualized with a Cy5-labeled peptide that binds to a specific extracytoplasmic site common to both proteins. Single-molecule trajectories were used to compute a radial distribution of displacements, yielding average diffusion coefficients equal to 0.22 (GPI-linked I-E(k)) and 0.18 microm(2)/s (native I-E(k)). The relative diffusion of pairs of proteins was also studied for intermolecular separations in the range 0.3-1.0 microm, to distinguish between free diffusion of a protein molecule and diffusion of proteins restricted to a rapidly diffusing small domain. Both analyses show that motion is predominantly Brownian. This study finds no strong evidence for significant confinement of either GPI-linked or native I-E(k) in the plasma membrane of CHO cells.

Full Text

The Full Text of this article is available as a PDF (394K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anderson RG. The caveolae membrane system. Annu Rev Biochem. 1998;67:199–225. [PubMed]
  • Anderson HA, Hiltbold EM, Roche PA. Concentration of MHC class II molecules in lipid rafts facilitates antigen presentation. Nat Immunol. 2000 Aug;1(2):156–162. [PubMed]
  • Berk DA, Hochmuth RM. Lateral mobility of integral proteins in red blood cell tethers. Biophys J. 1992 Jan;61(1):9–18. [PMC free article] [PubMed]
  • Brown DA, London E. Functions of lipid rafts in biological membranes. Annu Rev Cell Dev Biol. 1998;14:111–136. [PubMed]
  • Brown DA, London E. Structure and function of sphingolipid- and cholesterol-rich membrane rafts. J Biol Chem. 2000 Jun 9;275(23):17221–17224. [PubMed]
  • Dietrich C, Volovyk ZN, Levi M, Thompson NL, Jacobson K. Partitioning of Thy-1, GM1, and cross-linked phospholipid analogs into lipid rafts reconstituted in supported model membrane monolayers. Proc Natl Acad Sci U S A. 2001 Sep 11;98(19):10642–10647. [PMC free article] [PubMed]
  • Dietrich Christian, Yang Bing, Fujiwara Takahiro, Kusumi Akihiro, Jacobson Ken. Relationship of lipid rafts to transient confinement zones detected by single particle tracking. Biophys J. 2002 Jan;82(1 Pt 1):274–284. [PMC free article] [PubMed]
  • Edidin M, Stroynowski I. Differences between the lateral organization of conventional and inositol phospholipid-anchored membrane proteins. A further definition of micrometer scale membrane domains. J Cell Biol. 1991 Mar;112(6):1143–1150. [PMC free article] [PubMed]
  • Faivre-Sarrailh C, Gauthier F, Denisenko-Nehrbass N, Le Bivic A, Rougon G, Girault JA. The glycosylphosphatidyl inositol-anchored adhesion molecule F3/contactin is required for surface transport of paranodin/contactin-associated protein (caspr). J Cell Biol. 2000 Apr 17;149(2):491–502. [PMC free article] [PubMed]
  • Feder TJ, Brust-Mascher I, Slattery JP, Baird B, Webb WW. Constrained diffusion or immobile fraction on cell surfaces: a new interpretation. Biophys J. 1996 Jun;70(6):2767–2773. [PMC free article] [PubMed]
  • Ghosh RN, Webb WW. Automated detection and tracking of individual and clustered cell surface low density lipoprotein receptor molecules. Biophys J. 1994 May;66(5):1301–1318. [PMC free article] [PubMed]
  • Griffith IJ, Ghogawala Z, Nabavi N, Golan DE, Myer A, McKean DJ, Glimcher LH. Cytoplasmic domain affects membrane expression and function of an Ia molecule. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4847–4851. [PMC free article] [PubMed]
  • Hiscox Stephen, Hallett Maurice B, Morgan B Paul, van den Berg Carmen W. GPI-anchored GFP signals Ca2+ but is homogeneously distributed on the cell surface. Biochem Biophys Res Commun. 2002 May 3;293(2):714–721. [PubMed]
  • Huby RD, Dearman RJ, Kimber I. Intracellular phosphotyrosine induction by major histocompatibility complex class II requires co-aggregation with membrane rafts. J Biol Chem. 1999 Aug 6;274(32):22591–22596. [PubMed]
  • Huby RD, Weiss A, Ley SC. Nocodazole inhibits signal transduction by the T cell antigen receptor. J Biol Chem. 1998 May 15;273(20):12024–12031. [PubMed]
  • Jacobson K, Ishihara A, Inman R. Lateral diffusion of proteins in membranes. Annu Rev Physiol. 1987;49:163–175. [PubMed]
  • Kusumi A, Sako Y, Yamamoto M. Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells. Biophys J. 1993 Nov;65(5):2021–2040. [PMC free article] [PubMed]
  • Marshall KW, Wilson KJ, Liang J, Woods A, Zaller D, Rothbard JB. Prediction of peptide affinity to HLA DRB1*0401. J Immunol. 1995 Jun 1;154(11):5927–5933. [PubMed]
  • Munnelly HM, Brady CJ, Hagen GM, Wade WF, Roess DA, Barisas BG. Rotational and lateral dynamics of I-A(k) molecules expressing cytoplasmic truncations. Int Immunol. 2000 Sep;12(9):1319–1328. [PubMed]
  • Poo M, Cone RA. Lateral diffusion of rhodopsin in the photoreceptor membrane. Nature. 1974 Feb 15;247(5441):438–441. [PubMed]
  • Pralle A, Keller P, Florin EL, Simons K, Hörber JK. Sphingolipid-cholesterol rafts diffuse as small entities in the plasma membrane of mammalian cells. J Cell Biol. 2000 Mar 6;148(5):997–1008. [PMC free article] [PubMed]
  • Qian H, Sheetz MP, Elson EL. Single particle tracking. Analysis of diffusion and flow in two-dimensional systems. Biophys J. 1991 Oct;60(4):910–921. [PMC free article] [PubMed]
  • Rabinowitz JD, Vrljic M, Kasson PM, Liang MN, Busch R, Boniface JJ, Davis MM, McConnell HM. Formation of a highly peptide-receptive state of class II MHC. Immunity. 1998 Nov;9(5):699–709. [PubMed]
  • Reay PA, Kantor RM, Davis MM. Use of global amino acid replacements to define the requirements for MHC binding and T cell recognition of moth cytochrome c (93-103). J Immunol. 1994 Apr 15;152(8):3946–3957. [PubMed]
  • Reay PA, Wettstein DA, Davis MM. pH dependence and exchange of high and low responder peptides binding to a class II MHC molecule. EMBO J. 1992 Aug;11(8):2829–2839. [PMC free article] [PubMed]
  • Rotsch C, Radmacher M. Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study. Biophys J. 2000 Jan;78(1):520–535. [PMC free article] [PubMed]
  • Sako Y, Kusumi A. Compartmentalized structure of the plasma membrane for receptor movements as revealed by a nanometer-level motion analysis. J Cell Biol. 1994 Jun;125(6):1251–1264. [PMC free article] [PubMed]
  • Saxton MJ. Lateral diffusion in an archipelago. Single-particle diffusion. Biophys J. 1993 Jun;64(6):1766–1780. [PMC free article] [PubMed]
  • Saxton MJ. Anomalous diffusion due to obstacles: a Monte Carlo study. Biophys J. 1994 Feb;66(2 Pt 1):394–401. [PMC free article] [PubMed]
  • Saxton MJ. Single-particle tracking: effects of corrals. Biophys J. 1995 Aug;69(2):389–398. [PMC free article] [PubMed]
  • Saxton MJ. Single-particle tracking: the distribution of diffusion coefficients. Biophys J. 1997 Apr;72(4):1744–1753. [PMC free article] [PubMed]
  • Saxton MJ, Jacobson K. Single-particle tracking: applications to membrane dynamics. Annu Rev Biophys Biomol Struct. 1997;26:373–399. [PubMed]
  • Schmitt L, Boniface JJ, Davis MM, McConnell HM. Kinetic isomers of a class II MHC-peptide complex. Biochemistry. 1998 Dec 15;37(50):17371–17380. [PubMed]
  • Schütz GJ, Kada G, Pastushenko VP, Schindler H. Properties of lipid microdomains in a muscle cell membrane visualized by single molecule microscopy. EMBO J. 2000 Mar 1;19(5):892–901. [PMC free article] [PubMed]
  • Schütz GJ, Schindler H, Schmidt T. Single-molecule microscopy on model membranes reveals anomalous diffusion. Biophys J. 1997 Aug;73(2):1073–1080. [PMC free article] [PubMed]
  • Sheets ED, Lee GM, Simson R, Jacobson K. Transient confinement of a glycosylphosphatidylinositol-anchored protein in the plasma membrane. Biochemistry. 1997 Oct 14;36(41):12449–12458. [PubMed]
  • Simons K, Toomre D. Lipid rafts and signal transduction. Nat Rev Mol Cell Biol. 2000 Oct;1(1):31–39. [PubMed]
  • Simson R, Sheets ED, Jacobson K. Detection of temporary lateral confinement of membrane proteins using single-particle tracking analysis. Biophys J. 1995 Sep;69(3):989–993. [PMC free article] [PubMed]
  • Simson R, Yang B, Moore SE, Doherty P, Walsh FS, Jacobson KA. Structural mosaicism on the submicron scale in the plasma membrane. Biophys J. 1998 Jan;74(1):297–308. [PMC free article] [PubMed]
  • Smith PR, Morrison IE, Wilson KM, Fernández N, Cherry RJ. Anomalous diffusion of major histocompatibility complex class I molecules on HeLa cells determined by single particle tracking. Biophys J. 1999 Jun;76(6):3331–3344. [PMC free article] [PubMed]
  • Stevenson BR, Begg DA. Concentration-dependent effects of cytochalasin D on tight junctions and actin filaments in MDCK epithelial cells. J Cell Sci. 1994 Mar;107(Pt 3):367–375. [PubMed]
  • Suzuki K, Sheetz MP. Binding of cross-linked glycosylphosphatidylinositol-anchored proteins to discrete actin-associated sites and cholesterol-dependent domains. Biophys J. 2001 Oct;81(4):2181–2189. [PMC free article] [PubMed]
  • Thomas JL, Holowka D, Baird B, Webb WW. Large-scale co-aggregation of fluorescent lipid probes with cell surface proteins. J Cell Biol. 1994 May;125(4):795–802. [PMC free article] [PubMed]
  • Vacchino JF, McConnell HM. Peptide binding to active class II MHC protein on the cell surface. J Immunol. 2001 Jun 1;166(11):6680–6685. [PubMed]
  • Varma R, Mayor S. GPI-anchored proteins are organized in submicron domains at the cell surface. Nature. 1998 Aug 20;394(6695):798–801. [PubMed]
  • Wade WF, Freed JH, Edidin M. Translational diffusion of class II major histocompatibility complex molecules is constrained by their cytoplasmic domains. J Cell Biol. 1989 Dec;109(6 Pt 2):3325–3331. [PMC free article] [PubMed]
  • Weiss S. Fluorescence spectroscopy of single biomolecules. Science. 1999 Mar 12;283(5408):1676–1683. [PubMed]
  • Wettstein DA, Boniface JJ, Reay PA, Schild H, Davis MM. Expression of a class II major histocompatibility complex (MHC) heterodimer in a lipid-linked form with enhanced peptide/soluble MHC complex formation at low pH. J Exp Med. 1991 Jul 1;174(1):219–228. [PMC free article] [PubMed]
  • Wilson KM, Morrison IE, Smith PR, Fernandez N, Cherry RJ. Single particle tracking of cell-surface HLA-DR molecules using R-phycoerythrin labeled monoclonal antibodies and fluorescence digital imaging. J Cell Sci. 1996 Aug;109(Pt 8):2101–2109. [PubMed]
  • Zhang F, Crise B, Su B, Hou Y, Rose JK, Bothwell A, Jacobson K. Lateral diffusion of membrane-spanning and glycosylphosphatidylinositol-linked proteins: toward establishing rules governing the lateral mobility of membrane proteins. J Cell Biol. 1991 Oct;115(1):75–84. [PMC free article] [PubMed]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...