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. 1985 Nov 1;231(3):517–522. doi: 10.1042/bj2310517

Caldesmon is a Ca2+-regulatory component of native smooth-muscle thin filaments.

S B Marston, W Lehman
PMCID: PMC1152781  PMID: 2934055

Abstract

Thin-filament preparations from four smooth muscle types (gizzard, stomach, trachea, aorta) all activate myosin MgATPase activity, are regulated by Ca2+, and contain actin, tropomyosin and a 120000-140000-Mr protein in the molar proportions 1:1/7:1/26. The 120000-140000-Mr protein from all sources is a potent inhibitor of actomyosin ATPase activity. Peptide-mapping and immunological evidence is presented showing that it is identical with caldesmon. Quantitative immunological data suggest that caldesmon is a component of all the thin filaments and that the thin-filament-bound caldesmon accounts for all the caldesmon in intact tissue. The myosin light-chain kinase content of thin-filament preparations was found to be negligible. We propose that caldesmon-based thin-filament Ca2+ regulation is a physiological mechanism in all smooth muscles.

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Selected References

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

  1. 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]
  2. Chacko S., Rosenfeld A. Regulation of actin-activated ATP hydrolysis by arterial myosin. Proc Natl Acad Sci U S A. 1982 Jan;79(2):292–296. doi: 10.1073/pnas.79.2.292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  4. Driska S., Hartshorne D. J. The contractile proteins of smooth muscle. Properties and components of a Ca2+-sensitive actomyosin from chicken gizzard. Arch Biochem Biophys. 1975 Mar;167(1):203–212. doi: 10.1016/0003-9861(75)90457-9. [DOI] [PubMed] [Google Scholar]
  5. Ebashi S. The Croonian lecture, 1979: Regulation of muscle contraction. Proc R Soc Lond B Biol Sci. 1980 Mar 21;207(1168):259–286. doi: 10.1098/rspb.1980.0024. [DOI] [PubMed] [Google Scholar]
  6. Engvall E. Enzyme immunoassay ELISA and EMIT. Methods Enzymol. 1980;70(A):419–439. doi: 10.1016/s0076-6879(80)70067-8. [DOI] [PubMed] [Google Scholar]
  7. Hartshorne D. J., Siemankowski R. F. Regulation of smooth muscle actomyosin. Annu Rev Physiol. 1981;43:519–530. doi: 10.1146/annurev.ph.43.030181.002511. [DOI] [PubMed] [Google Scholar]
  8. Heaslip R. J., Chacko S. Effects of Ca2+ and Mg2+ on the actomyosin adenosine-5'-triphosphatase of stably phosphorylated gizzard myosin. Biochemistry. 1985 May 21;24(11):2731–2736. doi: 10.1021/bi00332a020. [DOI] [PubMed] [Google Scholar]
  9. Kakiuchi R., Inui M., Morimoto K., Kanda K., Sobue K., Kakiuchi S. Caldesmon, a calmodulin-binding, F actin-interacting protein, is present in aorta, uterus and platelets. FEBS Lett. 1983 Apr 18;154(2):351–356. doi: 10.1016/0014-5793(83)80181-1. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Lehman W., Head J. F., Grant P. W. The stoichiometry and location of troponin I- and troponin C-like proteins in the myofibril of the bay scallop, Aequipecten irradians. Biochem J. 1980 May 1;187(2):447–456. doi: 10.1042/bj1870447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Marston S. B., Moody C., Smith C. Mechanisms of the regulation of myofibrillar function in vascular smooth muscle. Biochem Soc Trans. 1984 Dec;12(6):945–948. doi: 10.1042/bst0120945. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Marston S. B., Trevett R. M., Walters M. Calcium ion-regulated thin filaments from vascular smooth muscle. Biochem J. 1980 Feb 1;185(2):355–365. doi: 10.1042/bj1850355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ngai P. K., Carruthers C. A., Walsh M. P. Isolation of the native form of chicken gizzard myosin light-chain kinase. Biochem J. 1984 Mar 15;218(3):863–870. doi: 10.1042/bj2180863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ngai P. K., Walsh M. P. Detection of caldesmon in muscle and non-muscle tissues of the chicken using polyclonal antibodies. Biochem Biophys Res Commun. 1985 Mar 15;127(2):533–539. doi: 10.1016/s0006-291x(85)80192-3. [DOI] [PubMed] [Google Scholar]
  17. Owada M. K., Hakura A., Iida K., Yahara I., Sobue K., Kakiuchi S. Occurrence of caldesmon (a calmodulin-binding protein) in cultured cells: comparison of normal and transformed cells. Proc Natl Acad Sci U S A. 1984 May;81(10):3133–3137. doi: 10.1073/pnas.81.10.3133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sellers J. R., Pato M. D. The binding of smooth muscle myosin light chain kinase and phosphatases to actin and myosin. J Biol Chem. 1984 Jun 25;259(12):7740–7746. [PubMed] [Google Scholar]
  19. 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]
  20. Sobieszek A., Small J. V. Myosin-linked calcium regulation in vertebrate smooth muscle. J Mol Biol. 1976 Mar 25;102(1):75–92. doi: 10.1016/0022-2836(76)90074-7. [DOI] [PubMed] [Google Scholar]
  21. Sobieszek A., Small J. V. Regulation of the actin-myosin interaction in vertebrate smooth muscle: activation via a myosin light-chain kinase and the effect of tropomyosin. J Mol Biol. 1977 Jun 5;112(4):559–576. doi: 10.1016/s0022-2836(77)80164-2. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Stewart D. I., Golosinska K., Smillie L. B. Identification of a troponin-I like protein in platelet preparations as histone H2B. FEBS Lett. 1983 Jun 27;157(1):129–132. doi: 10.1016/0014-5793(83)81130-2. [DOI] [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. Tsang V. C., Wilson B. C., Peralta J. M. Quantitative, single-tube, kinetic-dependent enzyme-linked immunosorbent assay (k-ELISA). Methods Enzymol. 1983;92:391–403. doi: 10.1016/0076-6879(83)92033-5. [DOI] [PubMed] [Google Scholar]
  26. Wang K., Ash J. F., Singer S. J. Filamin, a new high-molecular-weight protein found in smooth muscle and non-muscle cells. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4483–4486. doi: 10.1073/pnas.72.11.4483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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