Skip to main content
SpringerLink
Log in

Organization and dynamics of lipids in bovine brain coated and uncoated vesicles

  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

Three characteristics have been demonstrated by the chemical analysis of bovine brain coated vesicles following removal of the coat proteins: a high protein content, a high cholesterol/lipid ratio and a high percentage of phosphatidylethanolamine amongst the phospholipids.

The study of lipid bilayer organization and dynamics has been performed using the fluorescent probes pyrene and parinaric acid (cis and trans). This has allowed the study of both lateral mobility and rotational motion in the lipid bilayer of the coated and uncoated vesicles.

Lateral mobility in the fluid phase of the lipid is slightly reduced by the presence of the clathrin coat, as indicated by the lower diffusion coefficient of pyrene in coated compared with uncoated vesicles.

At all temperatures from 6° to 30°C, solid-phase domains, probed by trans parinaric acid, coexist with fluid-phase domains in the lipid bilayer. The temperature dependence of the parinaric acid lifetimes and of their amplitudes strongly suggests that the solid phase domains decrease in size with temperature, both in coated and uncoated vesicles.

However, the difference in the value of the anisotropy at long times (r ∞), between coated and uncoated vesicles (a difference which is more pronounced for cis than for trans parinaric acid), indicates that the presence of the clathrin coat introduces disorder in the surrounding lipids, thus suggesting a possible role of the clathrin in the formation of the pits on the plasma membrane.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

CVs:

coated vesicles

UVs:

uncoated vesicles

TLC:

thin layer chromatography

DMSO:

dimethylsulfoxide

DPPC:

dipalmitoylphosphatidylcholine

cis Pna:

cis parinaric acid

α (9,11,13,15-cis-trans-trans-cis):

octadecatetraenoic acid

Trans Pna:

Trans parinaric acid

β (9,11,13,15-all-trans):

octadecatetraenoic acid

References

  • Altstiel L, Branton D (1983) Fusion of coated vesicles with lysosomes: measurement with a fluorescent assay. Cell 32: 921–929

    Google Scholar 

  • Ames BN, Dubin DT (1960) The role of polyamines in the neutralization of bacteriophages deoxyribonucleic acid. J Biol Chem 235: 769–775

    Google Scholar 

  • Dickson RB, Beguinot L, Hanover JA, Richert ND, Willingham MC, Pastan I (1983) Isolation and characterization of a highly enriched preparation of receptosomes (endosomes) from a human cell line. Proc Natl Acad Sci USA 80: 5335–5339

    Google Scholar 

  • Eichberg J, Whittaker VP, Dawson RMC (1964) Distribution of lipids in subcellular particles of guinea pig brain. Biochem J 92: 91–96

    Google Scholar 

  • Galla HJ, Hartmann W (1980) Excimer forming lipids in membrane research. Chem Phys Lipids 27: 199–219

    Google Scholar 

  • Galla HJ, Luisetti J (1980) Lateral and transversal diffusion and phase transitions in erythrocyte membranes, an excimer fluorescence study. Biochim Biophys Acta 596: 108–117

    Google Scholar 

  • Galla HJ, Sackmann E (1974) Lateral diffusion in the hydrophobic region of membranes: use of pyrene excimer as optical probes. Biochim Biophys Acta 339: 103–115

    Google Scholar 

  • Galla HJ, Hartmann W, Theilen U, Sackmann E (1979) On two dimensional passive random walk in lipid bilayers and fluid pathways in biomembranes. J Membr Biol 48: 215–236

    Google Scholar 

  • Gallay J, Vincent M, de Paillerets C, Rogard M, Alfsen A (1981) Relationship between the activity of the 3β hydroxysteroid dehydrogenase from bovine adrenal cortex microsomes and membrane structure. J Biol Chem 256: 1235–1241

    Google Scholar 

  • Goldstein HL, Anderson RGW, Brown M (1979) Coated pits, coated vesicles and receptor mediated endocytosis. Nature 279: 679–685

    Google Scholar 

  • Hess HH, Rolde E (1964) Fluoremetric assay of sialic acid in brain gangliosides. J Biol Chem 239: 3215–3220

    Google Scholar 

  • Hess HH, Talheimer C (1965) Microassay of biochemical structural components in nervous tissues. Extraction and partition of lipids and assay of nucleic acids. J Neurochem 12: 193–204

    Google Scholar 

  • Heuser JE, Reese TS (1973) Evidence for recycling of synaptic vesicles membranes during transmitter release at the frog neuromuscular junction. J Cell Biol 57: 315–344

    Google Scholar 

  • Isenberg I, Small EW (1982) Exponential depression as a test of estimated decay parameters. J Chem Phys 77: 2799–2805

    Google Scholar 

  • Keen HJ, Willingham MC, Pastan IH (1979) Clathrin and coated vesicles proteins: immunological characterization. Cell 16: 303–312

    Google Scholar 

  • Kimmelman D, Tecoma ES, Wolber PK, Hudson BS, Wickner WT, Simoni RD (1979) Protein-lipid interactions. Studies of the M13 coat protein in dimyristoylphosphatidylcholine vesicles using parinaric acid. Biochemistry 18: 5874–5880

    Google Scholar 

  • Markwek MAK, Haas SM, Bieber LL, Talbert NE (1978) A modification of the Lowry procedure to simplify protein determination in membranes and lipoprotein samples. Anal Biochem 87: 206–210

    Google Scholar 

  • McGookey DJ, Fagerberg K, Anderson RGW (1983) Filipin-cholesterol complexes form in uncoated vesicles membrane derived from coated vesicles during receptor mediated endocytosis of low density lipoproteins. J Cell Biol 96: 1273–1278

    Google Scholar 

  • Montesano R, Perrelet A, Vassali P, Orci L (1980) Absence of Filipin-cholesterol complexes from large coated pits of cultured cells. Proc Natl Acad Sci USA 76: 6391–6395

    Google Scholar 

  • Nandi PK, Irace G, Van Jaarsveld PP, Lippoldt RE, Edelhoch H (1982) Instability of coated vesicles in concentrated sucrose solutions. Proc Natl Acad Sci USA 79: 5881–5885

    Google Scholar 

  • Norton WT, Autiliot LA (1966) The lipid composition of purified bovine brain myelin. J Neurochem 13: 213–217

    Google Scholar 

  • Nossal R, Weiss GH, Nandi PK, Lippoldt RE, Edelhoch H (1983) Sizes and mass distribution of clathrin coated vesicles from bovine brain. Arch Biochem Biophys 226: 593–603

    Google Scholar 

  • Ott P, Bingeli Y, Brodbeck Y (1982) A rapid and sensitive assay for determination of cholesterol in membrane lipid extracts. Biochim Biophys Acta 685: 211–213

    Google Scholar 

  • Paillerets de C, Gallay J, Vincent M, Rogard M, Alfsen A (1981) Membrane lipid dynamics and enzymatic activity in bovine adrenal microsomes. Biochim Biophys Acta 664: 134–142

    Google Scholar 

  • Pearse BMF (1975) Coated vesicles from pig brain: purification and biochemical characterization. J Mol Biol 97: 93–98

    Google Scholar 

  • Pearse BMF (1976) Clathrin: a unique protein associated with intracellular transfer of membranes by coated vesicles. Proc Natl Acad Sci USA 73: 1255–1259

    Google Scholar 

  • Pearse BMF (1982) Coated vesicles from human placenta carry ferritin, transferrin and immunoglobulin G. Proc Natl Acad Sci USA 79: 451–455

    Google Scholar 

  • Pearse BMF, Bretscher MS (1981) Membrane recycling by coated vesicles. Annu Rev Biochem 50: 85–101

    Google Scholar 

  • Roth TF, Woods JW (1982) In: Differentiation and function in hematopoietic cell surfaces. AR Liss, New York, USA, pp 163–181

    Google Scholar 

  • Rothman JE, Bursztyn-Pettegrew H, Fine HE (1980) Coated vesicles transport newly synthesized membrane glycoproteins from endoplasmic reticulum to plasma membrane in two successive stages. J Cell Biol 86: 162–171

    Google Scholar 

  • Salisbury JL, Condeelis JS, Satir P (1980) The role of coated vesicles microfilaments and calmodulin in receptor mediated endocytosis by cultured B lymphoblastoid cells. J Cell Biol 87: 132–141

    Google Scholar 

  • Sklar LA, Hudson BS, Petersen M, Diamond J (1977a) Conjugated polyene fatty acids as fluorescent probes: spectroscopic characterization. Biochemistry 16: 813–819

    Google Scholar 

  • Sklar LA, Hudson BS, Simoni RD (1977b) Conjugated polyene fatty acids as fluorescent probes: synthetic phospholipid membranes studies. Biochemistry 16: 819–828

    Google Scholar 

  • Tecoma ES, Sklar LA, Simoni RD, Hudson BS (1977) Conjugated polyene fatty acids as fluorescent probes: Biosynthetic incorporation of parinaric acid by Escherichia coli and studies of phase transitions. Biochemistry 16: 829–835

    Google Scholar 

  • Trewehella MA, Collins FD (1973) Nature of the fatty acyl chains in different phospholipids. Biochim Biophys Acta 296: 51–61

    Google Scholar 

  • Unanue ER, Ungewickell E, Branton D (1981) Clathrin membrane interaction. Cell 26: 439–446

    Google Scholar 

  • Vanderkooi JM, Collis JB (1974) Pyrene: a probe of lateral diffusion in the hydrophobic region of membranes. Biochemistry 13: 4000–4006

    Google Scholar 

  • Vincent M, de Foresta B, Gallay J, Alfsen A (1982) Nanosecond fluorescence anisotropy decays of n-(9-anthroyloxy)fatty acids in dipalmitoylcholine vesicles with regard to isotropic solvents. Biochemistry 21: 708–716

    Google Scholar 

  • Wolber PK, Hudson BS (1981) Fluorescence lifetime and time resolved polarization anisotropy studies of acyl chains order and dynamics in lipid bilayers. Biochemistry 20: 2800–2810

    Google Scholar 

  • Woodward MP, Roth RF (1978) Coated vesicles characterization, selective dissociation and reassembly. Proc Natl Acad Sci USA 75: 4394–4398

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. E.L.M.-E.R. 64 C.N.R.S., 45 rue des Saints Pères, F-75270, Paris, France

    A. Alfsen, C. de Paillerets, K. Prasad, P. K. Nandi & R. E. Lippoldt

  2. C.E.B., NIADDK, NIH, 20205, Bethesda, MD, USA

    H. Edelhoch

Authors
  1. A. Alfsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. de Paillerets
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K. Prasad
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. K. Nandi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R. E. Lippoldt
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H. Edelhoch
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Alfsen, A., de Paillerets, C., Prasad, K. et al. Organization and dynamics of lipids in bovine brain coated and uncoated vesicles. Eur Biophys J 11, 129–136 (1984). https://doi.org/10.1007/BF00276628

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00276628

Key words

Search

Navigation