Binding of dynamin mutants to lipid tubules
Mutants of dynamin were not only assayed for their conformation on lipid tubules in the presence of GTPgS and GTP but using a spin assay6 the amount that bound to the lipid tubules was also assayed (Fig. 3s). In this representative Coomassie gel all the mutants bind to the tubules with similar efficiencies.
1. Sever, S., Muhlberg, A. B. & Schmid, S. L. Impairment of dynamin's GAP domain stimulates receptor-mediated endocytosis. Nature 398, 481-486 (1999).
2. Sever, S., Damke, H. & Schmid, S. L. Dynamin:GTP controls the formation of constricted coated pits, the rate limiting step in clathrin-mediated endocytosis. J Cell Biol 150, 1137-1148 (2000).
3. Muhlberg, A. B., Warnock, D. E. & Schmid, S. L. Domain structure and intramolecular regulation of dynamin GTPase. EMBO J. 16, 6676-6683 (1998).
4. Okamoto, P. M., Tripet, B., Litowski, J., Hodges, R. S. & Vallee, R. B. Multiple distinct coiled-coils are involved in dynamin self-assembly. J Biol Chem 274, 10277-10286 (1999).
5. Herskovits, J. S., Burgess, C. C., Obar, R. A. & Vallee, R. B. Effects of mutant rat dynamin on endocytosis. J. Cell Biol. 122, 565-578 (1993).
6. Owen, D. J. et al. Crystal structure of the Amphiphysin-2 SH3 domain and its role in prevention of dynamin ring formation. EMBO J. 17, 5273-5285 (1998).
7. Binns, D. D. et al. The mechanism of GTP hydrolysis by dynamin II: a transient kinetic study. Biochemistry 39, 7188-7196 (2000).
8. Hinshaw, J. E. & Schmid, S. L. Dynamin self-assembles into rings suggesting a mechanism for coated vesicle budding. Nature 374, 190-192 (1995).
9. Warnock, D. E., HInshaw, J. E. & Schmid, S. L. Dynamin self-assembly stimulates its GTPase activity. J. Biol. Chem. 271, 22310-22314 (1996).