Summary
We have sequenced a cloned cDNA from a chicken gizzard library which encodes a short version of caldesmon with the properties expected for the non-muscle protein. The predicted protein is 524 amino acids long with a molecular mass of 60174 daltons. Expression in bacteria produces a protein with an apparent molecular weight on SDS gels of 75–77000, that comigrates with human platelet caldesmon. The expressed protein binds to F-actin and is retained on calmodulin-Sepharose in the presence, but not absence of Ca2+. This non-muscle caldesmon isoform is identical to the smooth muscle protein at the N- and C-terminal ends, but is missing 232 amino acids from the centre. This central segment is thought to be helical, suggesting that non-muscle caldesmon is approximately 35 nm shorter, about 40 nm overall length, than smooth muscle caldesmon.
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References
Ball, E. H. &Kovala, T. (1988) Mapping of caldesmon: relationship between the high and low molecular weight forms.Biochemistry 27, 6093–8.
Bretscher, A. &Lynch, W. (1985) Identification and localization of immunoreactive forms of caldesmon in smooth and non-muscle cells: a comparison with the distributions of tropomyosin and alpha-actinin.J. Cell Biol. 100, 1656–63.
Bryan, J. (1989) Caldesmon, acidic amino acids and molecular weight determinations.J. Muscle Res. Cell Motil. 10, 95–6.
Bryan, J. (1990) Caldesmon: fragments, sequence and domain mapping.Ann. N. Y. Acad. Sci. 599, 100–10.
Bryan, J., Imai, M., Lee, R., Moore, P., Cook, R. G. &Lin, W-G. (1989) Cloning and expression of a smooth muscle caldesmon.J. Biol. Chem. 264, 13873–9.
Chalovich, J.M. (1988) Caldesmon and thin-filament regulation of muscle contraction.Cell Biophys. 12, 73–85.
Dingus, J., Hwo, S. &Bryan, J. (1986) Identification by monoclonal antibodies and characterization of human platelet caldesmon.J. Cell Biol. 102, 1748–57.
Fechheimer, M., Murdock, D., Carney, M. &Glover, C. V. C. (1991) Isolation and sequencing of cDNA clones encoding theDictyostelium discoideum 30000 dalton actin bundling protein.J. Biol. Chem. 266, 2883–9.
Fujii, T., Ozawa, J., Ogoma, Y. &Kondo, Y. (1988) Interaction between chicken gizzard caldesmon and tropomyosin.J. Biochem. (Tokyo) 104, 734–7.
Furst, D. O., Cross, R. A., De Mey, J. &Small, J. V. (1986) Caldesmon is an elongated, flexible molecule localized in the actomyosin domains of smooth muscle.EMBO J. 5, 251–7.
Graceffa, P. (1987) Evidence for interaction between smooth muscle tropomyosin and caldesmon.FEBS Lett. 218, 139–42.
Graceffa, P., Wang, C. -L. A. &Stafford, W. F. (1988) Caldesmon. Molecular weight and subunit composition by analytical ultracentrifugation.J. Biol. Chem. 263, 14196–202.
Hayashi, K., Kanda, K., Kimizuka, F., Kato, I &Sobue, K. (1989) Primary structure and functional expression of h-caldesmon complementary DNA.Biochem. Biophys. Res. Commun. 164, 503–11.
Hemric, M. E. &Chalovich, J. M. (1988) Effect of caldesmon on the ATPase activity and the binding of smooth and skeletal myosin subfragments to actin.J. Biol. Chem. 263, 1878–85.
Horiuchi, K. Y. &Chacko, S. (1988) Interaction between caldesmon and tropomyosin in the presence and absence of smooth muscle actin.Biochem. 27, 8388–93.
Ikebe, M. &Reardon, S. (1988) Binding of caldesmon to smooth muscle myosin.J. Biol. Chem. 263, 3055–8.
Koji-Owada, M., Hakura, A., Iida, K., Yahara, I., Sobue, K. &Kakiuchi, S. (1984)Proc. Natl. Acad. Sci. USA 81, 3133–7.
Kozak, M. (1988) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs.Nucleic Acids Res. 15, 8125–48.
Lash, J. A., Sellers, J. R. &Hathaway, D. R. (1986) The effects of caldesmon on smooth muscle heavy actomeromyosin ATPase activity and binding of heavy meromyosin to actin.J. Biol. Chem. 261, 16155–60.
Mabuchi, K. &Wang, C.-L. A (1991) Electron microscopic studies of chicken gizzard caldesmon and its complex with calmodulin.J. Muscle Res. Cell Motil. 12, 145–51.
Marston, S. B. &Smith C. W. J. (1985) The thin filaments of smooth muscles.J. Muscle Res. Cell Motil. 6, 669–708.
Nagai, K &Thogersen, H. V. (1987) Synthesis and sequence-specific proteolysis of hybrid proteins produced inEscherichia coli.Meth. Enzymol. 153, 461–81.
Novy, R. E., Lin, J. L -C. &Lin J. J -C. (1990) Isolation of a cDNA encoding a low Mr human caldesmon isoform.J. Cell Biol. 111, 163a.
Riseman, V. M., Lynch, W. P., Nefsky, B. &Bretscher, A. (1989) The calmodulin and F-actin binding sites of smooth muscle caldesmon lie in the carboxyl-terminal domain whereas the molecular weight heterogeneity lies in the middle of the molecule.J. Biol. Chem. 264, 2869–75.
Sobue, K., Muramoto, Y., Fujita, M. &Kakiuchi, S. (1981) Purification of a calmodulin-binding protein from chicken gizzard that interacts with F-actin.Proc. Natl Acad. Sci. USA.78, 5652–5
Stafford, W. F., Jancso, A. &Graceffa, P. (1990) Caldesmon from rabbit liver: molecular weight and length by analytical ultracentrifugation.Arch. Biochem. Biophys. 281, 66–9.
Ueki, N., Sobue, K., Kanda, K., Hada-T. &Higashino, K. (1987) Expression of high and low molecular weight caldesmons during phenotypic modulation of smooth muscle cells.Proc. Natl Acad. Sci. USA. 84, 9049–53.
Wang, C.-L. A., Wang, L.-W. C., Xu, S., Lu, R. C., Saavedra-Alanis, V. &Bryan, J. (1991) Localization of the calmodulin- and Actin-binding sites of caldesmon.J. Biol. Chem. (in press).
Yamakita, Y., Yamashiro, S. &Matsumura, F. (1990) Microinjection of non-muscle and smooth muscle caldesmon into fibroblasts and muscle cells.J. Cell Biol. 111, 2487–98.
Yamashiro-Matsumura, S., Ishikawa, R. &Matsumura, F. Purification and characterization of 83 kDa non-muscle caldesmon from cultured rat cells: changes in its expression upon L6 myogenesis. (1988)Protoplasma (Suppl) 2, 9–21.
Yamashiro-Matsumura, S. &Matsumura, F. (1988) Characterization of 83-kilodalton non-muscle caldesmon from cultured rat cells: stimulation of actin binding of nonmuscle tropomyosin and periodic localization along microfilaments like tropomyosin.J. Cell Biol. 106, 1973–83.
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Bryan, J., Lee, R. Sequence of an avian non-muscle caldesmon. J Muscle Res Cell Motil 12, 372–375 (1991). https://doi.org/10.1007/BF01738592
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DOI: https://doi.org/10.1007/BF01738592