Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1986 Feb;5(2):251–257. doi: 10.1002/j.1460-2075.1986.tb04206.x

Caldesmon is an elongated, flexible molecule localized in the actomyosin domains of smooth muscle.

D O Fürst, R A Cross, J De Mey, J V Small
PMCID: PMC1166726  PMID: 3709514

Abstract

A rapid purification procedure has been developed for the isolation of caldesmon from hog stomach smooth muscle utilizing a KI extract of washed myofibrils as source material. On SDS-PAGE this mammalian caldesmon showed a closely-spaced doublet around 155 kd. By low-angle rotary shadowing caldesmon was shown to be an elongated, highly flexible molecule which tends to form end-to-end dimers that are structurally very similar to filamin. When added to F-actin solutions caldesmon increased the high-shear viscosity considerably, but by an extent that depended on sample preparation. The effect was shown to be due to caldesmon and not to a trace contaminant by its full reversibility after addition of a monospecific caldesmon antibody. Recent investigations have shown that in smooth muscle two structurally distinct domains can be distinguished: an actomyosin domain and an actin-intermediate filament domain. Immunocytochemistry of ultrathin sections of smooth muscle at the light and electron microscope level revealed that caldesmon is present in the actomyosin domain. Caldesmon is thus a potential regulator of the actomyosin system in smooth muscle.

Full text

PDF
251

Images in this article

251Fig. 1.
on p.251
252Fig. 2.
on p.252
254Fig. 4.
on p.254
255Fig. 5.
on p.255

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bretscher A., Lynch W. Identification and localization of immunoreactive forms of caldesmon in smooth and nonmuscle cells: a comparison with the distributions of tropomyosin and alpha-actinin. J Cell Biol. 1985 May;100(5):1656–1663. doi: 10.1083/jcb.100.5.1656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bretscher A. Smooth muscle caldesmon. Rapid purification and F-actin cross-linking properties. J Biol Chem. 1984 Oct 25;259(20):12873–12880. [PubMed] [Google Scholar]
  3. Burridge K., Connell L. A new protein of adhesion plaques and ruffling membranes. J Cell Biol. 1983 Aug;97(2):359–367. doi: 10.1083/jcb.97.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Castellani L., Offer G., Elliott A., O'Brien E. J. Structure of filamin and the F-actin-heavy merofilamin complex. J Muscle Res Cell Motil. 1981 Jun;2(2):193–202. doi: 10.1007/BF00711869. [DOI] [PubMed] [Google Scholar]
  5. De Mey J., Moeremans M., Geuens G., Nuydens R., De Brabander M. High resolution light and electron microscopic localization of tubulin with the IGS (immuno gold staining) method. Cell Biol Int Rep. 1981 Sep;5(9):889–899. doi: 10.1016/0309-1651(81)90204-6. [DOI] [PubMed] [Google Scholar]
  6. Ebashi S., Ebashi F. Alpha-actinin, a new structural protein from striated muscle. I. Preparation and action on actomyosinàtp interaction. J Biochem. 1965 Jul;58(1):7–12. doi: 10.1093/oxfordjournals.jbchem.a128167. [DOI] [PubMed] [Google Scholar]
  7. Elliott A., Offer G. Shape and flexibility of the myosin molecule. J Mol Biol. 1978 Aug 25;123(4):505–519. doi: 10.1016/0022-2836(78)90204-8. [DOI] [PubMed] [Google Scholar]
  8. Fornstedt N. Affinity chromatographic studies on antigen-antibody dissociation. FEBS Lett. 1984 Nov 19;177(2):195–199. doi: 10.1016/0014-5793(84)81282-x. [DOI] [PubMed] [Google Scholar]
  9. Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. doi: 10.1016/0092-8674(79)90368-4. [DOI] [PubMed] [Google Scholar]
  10. Gopalakrishna R., Anderson W. B. Ca2+-induced hydrophobic site on calmodulin: application for purification of calmodulin by phenyl-Sepharose affinity chromatography. Biochem Biophys Res Commun. 1982 Jan 29;104(2):830–836. doi: 10.1016/0006-291x(82)90712-4. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Marston S. B. The regulation of smooth muscle contractile proteins. Prog Biophys Mol Biol. 1983;41(1):1–41. doi: 10.1016/0079-6107(83)90024-x. [DOI] [PubMed] [Google Scholar]
  13. Matsudaira P. T., Burgess D. R. SDS microslab linear gradient polyacrylamide gel electrophoresis. Anal Biochem. 1978 Jul 1;87(2):386–396. doi: 10.1016/0003-2697(78)90688-7. [DOI] [PubMed] [Google Scholar]
  14. Moeremans M., Daneels G., Van Dijck A., Langanger G., De Mey J. Sensitive visualization of antigen-antibody reactions in dot and blot immune overlay assays with immunogold and immunogold/silver staining. J Immunol Methods. 1984 Nov 30;74(2):353–360. doi: 10.1016/0022-1759(84)90303-x. [DOI] [PubMed] [Google Scholar]
  15. Ngai P. K., Walsh M. P. Inhibition of smooth muscle actin-activated myosin Mg2+-ATPase activity by caldesmon. J Biol Chem. 1984 Nov 25;259(22):13656–13659. [PubMed] [Google Scholar]
  16. Shotton D. M., Burke B. E., Branton D. The molecular structure of human erythrocyte spectrin. Biophysical and electron microscopic studies. J Mol Biol. 1979 Jun 25;131(2):303–329. doi: 10.1016/0022-2836(79)90078-0. [DOI] [PubMed] [Google Scholar]
  17. Small J. V., Fürst D. O., De Mey J. Localization of filamin in smooth muscle. J Cell Biol. 1986 Jan;102(1):210–220. doi: 10.1083/jcb.102.1.210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith C. W., Marston S. B. Disassembly and reconstitution of the Ca2+-sensitive thin filaments of vascular smooth muscle. FEBS Lett. 1985 May 6;184(1):115–119. doi: 10.1016/0014-5793(85)80665-7. [DOI] [PubMed] [Google Scholar]
  19. Sobieszek A., Bremel R. D. Preparation and properties of vertebrate smooth-muscle myofibrils and actomyosin. Eur J Biochem. 1975 Jun 16;55(1):49–60. doi: 10.1111/j.1432-1033.1975.tb02137.x. [DOI] [PubMed] [Google Scholar]
  20. Sobue K., Morimoto K., Kanda K., Maruyama K., Kakiuchi S. Reconstitution of Ca2+-sensitive gelation of actin filaments with filamin, caldesmon and calmodulin. FEBS Lett. 1982 Feb 22;138(2):289–292. doi: 10.1016/0014-5793(82)80463-8. [DOI] [PubMed] [Google Scholar]
  21. Sobue K., Muramoto Y., Fujita M., Kakiuchi S. Purification of a calmodulin-binding protein from chicken gizzard that interacts with F-actin. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5652–5655. doi: 10.1073/pnas.78.9.5652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sobue K., Takahashi K., Tanaka T., Kanda K., Ashino N., Kakiuchi S., Maruyama K. Crosslinking of actin filaments is caused by caldesmon aggregates, but not by its dimers. FEBS Lett. 1985 Mar 11;182(1):201–204. doi: 10.1016/0014-5793(85)81184-4. [DOI] [PubMed] [Google Scholar]
  23. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tyler J. M., Anderson J. M., Branton D. Structural comparison of several actin-binding macromolecules. J Cell Biol. 1980 May;85(2):489–495. doi: 10.1083/jcb.85.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wang K. Filamin, a new high-molecular-weight protein found in smooth muscle and nonmuscle cells. Purification and properties of chicken gizzard filamin. Biochemistry. 1977 May 3;16(9):1857–1865. doi: 10.1021/bi00628a015. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES