Advertisement

Liposomes pp 531-542 | Cite as

Use of Liposomes to Study Vesicular Transport

  • Kohji TakeiEmail author
  • Hiroshi Yamada
  • Tadashi Abe
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 606)

Abstract

Liposomes have been utilized for variety of membrane transport studies including clathrin-mediated endocytosis. Here we introduce clathrin-coated structures that are generated on large unilamellar liposomes by incubating with clathrin coat proteins. Large unilamellar liposomes are also used to reconstitute vesicle formation in a cell-free system, and the vesicle formation can be quantified by using dynamic light scattering (DLS). Furthermore, phagocytosis assay using liposome-conjugated styrene beads is demonstrated.

Key words

Vesicular transport Membrane traffic Endocytosis Phagocytosis 

Notes

Acknowledgement

The work was supported in part by grants from the Ministry of Education, Science, Sports and Culture of Japan.

References

  1. 1.
    Balch WE, Dunphy WG, Braell WA, Rothman JE (1984) Reconstitution of the transport of protein between successive compartments of the Golgi measured by the coupled incorporation of N-acetylglucosamine. Cell 39:405-416CrossRefPubMedGoogle Scholar
  2. 2.
    Orci L, Glick BS, Rothman JE (1986) A new type of coated vesicular carrier that appears not to contain clathrin: its possible role in protein transport within the Golgi stack. Cell 46:171-184CrossRefPubMedGoogle Scholar
  3. 3.
    Barlowe C, Orci L, Yeung T, Hosobuchi M, Hamamoto S, Salama N, Rexach MF, Ravazzola M, Amherdt M, Schekman R (1994) COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum. Cell 77:895-907CrossRefPubMedGoogle Scholar
  4. 4.
    Spang A, Matsuoka K, Hamamoto S, Schekman R, Orci L (1998) Free in PMCCoatomer, Arf1p, and nucleotide are required to bud coat protein complex I-coated vesicles from large synthetic liposomes. Proc Natl Acad Sci U S A 95:11199-11204CrossRefPubMedGoogle Scholar
  5. 5.
    Matsuoka K, Schekman R (2000) The use of liposomes to study COPII- and COPI-coated vesicle formation and membrane protein sorting. Methods 20:417-428CrossRefPubMedGoogle Scholar
  6. 6.
    Takei K, Haucke V, Slepnev V, Farsad K, Salazar M, Chen H et al (1998) Generation of coated intermediates of clathrin-mediated endocytosis on protein-free liposomes. Cell 94:131-141CrossRefPubMedGoogle Scholar
  7. 7.
    Kinuta M, Yamada H, Abe T, Watanabe M, Li S-I, Kamitani A et al (2002) Phosphatidylinositol 4, 5-bisphosphate stimulates vesicle formation from liposomes by brain cytosol. Proc Natl Acad Sci U S A 99:2842-2847CrossRefPubMedGoogle Scholar
  8. 8.
    Kinuta M, Takei K (2002) Utilization of liposomes in vesicular transport studies. Cell Struct Funct 27:63-69CrossRefPubMedGoogle Scholar
  9. 9.
    Greenberg S, Grinstein S (2002) Phagocytosis and innate immunity. Curr Opin Immunol 14:136-145CrossRefPubMedGoogle Scholar
  10. 10.
    Shiratsuchi A, Umeda M, Ohba Y, Nakanishi Y (1997) Recognition of phosphatidylserine on the surface of apoptotic spermato­­genic cells and subsequent phagocytosis by Sertoli cells of the rat. J Biol Chem 272:2354-2358CrossRefPubMedGoogle Scholar
  11. 11.
    Yamada H, Ohashi E, Abe T, Kusumi N, Li SA, Yoshida Y et al (2007) Amphiphysin 1 is important for actin polymerization during phagocytosis. Mol Biol Cell 18:4669-4680CrossRefPubMedGoogle Scholar
  12. 12.
    Maycox PR, Link E, Reetz A, Morris SA, Jahn R (1992) Clathrin-coated vesicles in nervous tissue are involved primarily in synaptic vesicle recycling. J Cell Biol 118:1379-1388CrossRefPubMedGoogle Scholar
  13. 13.
    Berne BJ, Pecora R (1976) Dynamic light scattering with applications to chemistry, biology and physics. John Wiley and Sons, New YorkGoogle Scholar
  14. 14.
    Otsuka A, Abe T, Watanabe M, Yagisawa H, Takei K, Yamada H (2009) Dynamin 2 is required for actin assembly in phagocytosis in Sertoli cells. Biochem Biophys Res Commun 378:478-482CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of NeuroscienceOkayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesOkayamaJapan

Personalised recommendations