Abstract
Familial hypertrophic cardiomyopathy(FHC) is caused by missence mutations in β-myosin heavy chain or other various sarcomeric proteins. To elucidate the functional impact of FHC mutations in myosin heavy chain, we generated Dictyostelium discoideum myosin II mutants equivalent to human FHC mutations by site-directed mutagenesis, and characterized their molecular-basis motor function. The current mutants, i.e. R397Q, F506C, G575R, A699R, K703Q and K703W are equivalent to R403Q, F513C, G584R, G716R, R719Q and R719W FHC mutants respectively. We measured the molecular-basis force and the sliding velocity generated by these myosin mutants. The measurement revealed that the A699R, K703Q and K703W myosins exhibited the lowest level of force with their preserved actin-activated MgATPase activity. F506C mutant showed the least impairment of the motile and enzymatic activities. The motor function of R397Q and G575R myosins were classified as intermediate. These results suggest that ELC binding domain might be important for force production.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Tanigawa, G., et al. Cell 62, 991–998 (1990).
Geisterfer, L.A., et al. Cell 62, 999–1006 (1990).
Seidman, C.E. & Seidman, J.G. Mol. Biol. Med. 8, 159–166 (1991).
Thierfelder, L., et al. Cell 77, 701–712 (1994).
Watkins, H., et al. Nat. Genet. 11, 434–437 (1995).
Watkins, H., et al. N. Engl. J. Med. 332, 1058–1064 (1995).
Watkins, H., et al. Nat. Genet. 3, 333–337 (1993).
Bonne, G., et al. Nat Genet 11, 438–440 (1995).
Poetter, K., et al. Nature Genet 13, 63–69 (1996).
Rayment, I., Holden, H.M., Sellers, J.R., Fananapazir, L. & Epstein, N.D. Proc. Natl. Acad. Sci. USA 92, 3864–3868 (1995).
Fisher, A.J., et al. Biochemistry 34, 8960–8972 (1995).
Kunkel, T.A. Proc. Natl. Acad. Sci. USA 82, 488–492 (1985).
Kunkel, T.A., Roberts, J.D. & Zakour, R.A. Methods Enzymol., 367–382 (1987).
Howard, P.K., Ahern, K.G. & Firtel, R.A. Nucleic. Acids. Res. 16, 2613–2623 (1988).
Manstein, D.J., Titus, M.A., De Lozanne, A. & Spudich, J.A. Embo. J. 8, 923–932 (1989).
Ruppel, K.M., Egelhoff, T.T. & Spudich, J.A. Ann. N. Y. Acad. Sci., 147–155 (1990).
Spudich, J.A. & Watt, S. J. Biol. Chem., 4866–4871 (1971).
Miyata, H., et al. J. Biochem. 115, 644–647 (1994).
Kamimura, S. Appl. Opt. 26, 3425–3434 (1987).
Svoboda, K. & Block, S.M. Proc. Natl. Acad. Sci. USA 91, 11782–11786 (1994).
Kron, S.J. & Spudich, J.A. Proc. Natl. Acad. Sci. USA 83, 6272–6276 (1986).
Kodama, T., Fukui, K. & Kometani, K. J. Biochem.(Tokyo) 99, 1465–1472 (1986).
Smith, C.A. & Rayment, I. Biochemistry 35, 5404–5417 (1996).
Whittaker, M., et al. Nature 378, 748–753 (1995).
Uyeda, T.Q., Abramson, P.D. & Spudich, J.A. Proc. Natl. Acad. Sci. USA 93, 4459–4464 (1996).
Milligan, R.A. Proc. Natl. Acad. Sci. USA 93, 21–26 (1996).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Plenum Press, New York
About this chapter
Cite this chapter
Fujita, H., Sugiura, S., Momomura, Si., Sugi, H., Sutoh, K. (1998). Functional Characterization of Dictyostelium Discoideum Mutant Myosins Equivalent to Human Familial Hypertrophic Cardiomyopathy. In: Sugi, H., Pollack, G.H. (eds) Mechanisms of Work Production and Work Absorption in Muscle. Advances in Experimental Medicine and Biology, vol 453. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-6039-1_16
Download citation
DOI: https://doi.org/10.1007/978-1-4684-6039-1_16
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4684-6041-4
Online ISBN: 978-1-4684-6039-1
eBook Packages: Springer Book Archive