Methods to Study the Interactions of the Dynein Light Chains and Intermediate Chains

  • Kevin W.-H. Lo
  • K. Kevin Pfister
Part of the Methods in Molecular Biology™ book series (MIMB, volume 392)


The cargo-binding domain of the cytoplasmic dynein complex consists of an intermediate chain, a light-intermediate chain, and three families of light chains. These five subunits form the base of the dynein complex. Variations in the composition and interactions of these subunits play an important role for selecting a particular cargo and regulating dynein function. By using several complementary binding methods, we have investigated the protein-protein interaction in the cargo-binding domain of the cytoplasmic dynein.

Key Words

Cytoplasmic dynein motor protein microtubule cytoskeleton intermediate chain light chain protein-protein interaction 


  1. 1.
    Pfister, K.K., Fisher, E.M., Gibbons, I.R., Hays, T.S., Holzbaur, E.L., McIntosh, J.R., Porter, M.E., Schroer, T.A., Vaughan, K.T., Witman, G.B., King, S.M., and Vallee, R.B. (2005) Cytoplasmic dynein nomenclature. J. Cell Biol 171, 411–413.CrossRefPubMedGoogle Scholar
  2. 2.
    Paschal, B.M. and Vallee, R.B. (1987) Retrograde transport by the microtubule-associated protein MAP 1C. Nature 330, 181–183.CrossRefPubMedGoogle Scholar
  3. 3.
    Pfister, K.K., Shah, P.R., Hummerich, H., Russ, A., Cotton, J., Annuar, A.A., King, S.M., and Fisher, E.M.C. (2006) Genetic analysis of the cytoplasmic dynein subunit families. PLoS Genet. 2, 11–26.CrossRefGoogle Scholar
  4. 4.
    Vallee, R.B., Williams, J.C., Varma, D., and Barnhart, L.E. (2004) Dynein: an ancient motor protein involved in multiple modes of transport. J. Neurobiol. 58, 189–200.CrossRefPubMedGoogle Scholar
  5. 5.
    King, S.M. (2000) The dynein microtubule motor. Biochim. Biophys. Acta 1496, 60–75.CrossRefPubMedGoogle Scholar
  6. 6.
    Vale, R.D. (2003) The molecular motor toolbox for intracellular transport. Cell 112, 467–480.CrossRefPubMedGoogle Scholar
  7. 7.
    Gee, M.A., Heuser, J.E., and Vallee, R.B. (1997) An extended microtubule-binding structure within the dynein motor domain. Nature 390, 636–639.CrossRefPubMedGoogle Scholar
  8. 8.
    Vallee, R.B. and Sheetz, M.P. (1996) Targeting of motor proteins. Science 271, 1539–1544.CrossRefPubMedGoogle Scholar
  9. 9.
    Vaughan, K.T. and Vallee, R.B. (1995) Cytoplasmic dynein binds dynactin through a direct interaction between the intermediate chains and p150Glued. J. Cell Biol. 131, 1507–1516.CrossRefPubMedGoogle Scholar
  10. 10.
    Lo, K.W., Naisbitt, S., Fan, J.S., Sheng, M., and Zhang, M. (2001) The 8-kDa dynein light chain binds to its targets via a conserved (K/R)XTQT motif. J. Biol. Chem. 276, 14059–14066.CrossRefPubMedGoogle Scholar
  11. 11.
    Mok, Y.K., Lo, K.W., and Zhang, M. (2001) Structure of Tctex-1 and its interaction with cytoplasmic dynein intermediate chain. J. Biol. Chem. 276, 14067–14074.PubMedGoogle Scholar
  12. 12.
    Susalka, S.J., Nikulina, K., Salata, M.W., Vaughan, P.S., King, S.M., Vaughan, K.T., and Pfister, K.K. (2002) The roadblock light chain binds a novel region of the cytoplasmic dynein intermediate chain. J. Biol. Chem. 277, 32939–32946.CrossRefPubMedGoogle Scholar
  13. 13.
    Lo, K.W., Kan, H.M., and Pfister, K.K. (2006) Identification of a novel region of the cytoplasmic dynein intermediate chain important for dimerization in the absence of the light chains. J. Biol. Chem. 281, 9552–9559.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Kevin W.-H. Lo
    • 1
  • K. Kevin Pfister
    • 1
  1. 1.Department of Cell BiologyUniversity of Virginia School of MedicineCharlottesville

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