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. 1991 Nov 15;280(Pt 1):33–38. doi: 10.1042/bj2800033

Functional interrelationship between calponin and caldesmon.

R Makuch 1, K Birukov 1, V Shirinsky 1, R Dabrowska 1
PMCID: PMC1130595  PMID: 1835840

Abstract

Calponin and caldesmon, constituents of smooth-muscle thin filaments, are considered to be potential modulators of smooth-muscle contraction. Both of them interact with actin and inhibit ATPase activity of smooth- and skeletal-muscle actomyosin. Here we show that calponin and caldesmon could bind simultaneously to F-actin when used in subsaturating amounts, whereas each one used in excess caused displacement of the other from the complex with F-actin. Calponin was more effective than caldesmon in this competition: when F-actin was saturated with calponin the binding of caldesmon was eliminated almost completely, whereas even at high molar excess of caldesmon one-third of calponin (relative to the saturation level) always remained bound to actin. The inhibitory effects of low concentrations of calponin and caldesmon on skeletal-muscle actomyosin ATPase were additive, whereas the maximum inhibition of the ATPase attained at high concentration of each of them was practically unaffected by the other one. These data suggest that calponin and caldesmon cannot operate on the same thin filaments. CA(2+)-calmodulin competed with actin for calponin binding, and at high molar excess dissociated the calponin-actin complex and reversed the calponin-induced inhibition of actomyosin ATPase activity.

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Selected References

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  1. Abe M., Takahashi K., Hiwada K. Effect of calponin on actin-activated myosin ATPase activity. J Biochem. 1990 Nov;108(5):835–838. doi: 10.1093/oxfordjournals.jbchem.a123289. [DOI] [PubMed] [Google Scholar]
  2. Abougou J. C., Hagiwara M., Hachiya T., Terasawa M., Hidaka H., Hartshorne D. J. Phosphorylation of caldesmon. FEBS Lett. 1989 Nov 6;257(2):408–410. doi: 10.1016/0014-5793(89)81583-2. [DOI] [PubMed] [Google Scholar]
  3. Adelstein R. S., Eisenberg E. Regulation and kinetics of the actin-myosin-ATP interaction. Annu Rev Biochem. 1980;49:921–956. doi: 10.1146/annurev.bi.49.070180.004421. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Bryan J., Imai M., Lee R., Moore P., Cook R. G., Lin W. G. Cloning and expression of a smooth muscle caldesmon. J Biol Chem. 1989 Aug 15;264(23):13873–13879. [PubMed] [Google Scholar]
  7. Bárány M., Rokolya A., Bárány K. Absence of calponin phosphorylation in contracting or resting arterial smooth muscle. FEBS Lett. 1991 Feb 11;279(1):65–68. doi: 10.1016/0014-5793(91)80252-x. [DOI] [PubMed] [Google Scholar]
  8. Dabrowska R., Goch A., Gałazkiewicz B., Osińska H. The influence of caldesmon on ATPase activity of the skeletal muscle actomyosin and bundling of actin filaments. Biochim Biophys Acta. 1985 Sep 27;842(1):70–75. doi: 10.1016/0304-4165(85)90295-8. [DOI] [PubMed] [Google Scholar]
  9. Fujii T., Ozawa J., Ogoma Y., Kondo Y. Interaction between chicken gizzard caldesmon and tropomyosin. J Biochem. 1988 Nov;104(5):734–737. doi: 10.1093/oxfordjournals.jbchem.a122542. [DOI] [PubMed] [Google Scholar]
  10. Fürst D. O., Cross R. A., De Mey J., Small J. V. Caldesmon is an elongated, flexible molecule localized in the actomyosin domains of smooth muscle. EMBO J. 1986 Feb;5(2):251–257. doi: 10.1002/j.1460-2075.1986.tb04206.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gimona M., Herzog M., Vandekerckhove J., Small J. V. Smooth muscle specific expression of calponin. FEBS Lett. 1990 Nov 12;274(1-2):159–162. doi: 10.1016/0014-5793(90)81353-p. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Graceffa P. Evidence for interaction between smooth muscle tropomyosin and caldesmon. FEBS Lett. 1987 Jun 22;218(1):139–142. doi: 10.1016/0014-5793(87)81034-7. [DOI] [PubMed] [Google Scholar]
  14. Hemric M. E., Chalovich J. M. Characterization of caldesmon binding to myosin. J Biol Chem. 1990 Nov 15;265(32):19672–19678. [PMC free article] [PubMed] [Google Scholar]
  15. Horiuchi K. Y., Chacko S. Interaction between caldesmon and tropomyosin in the presence and absence of smooth muscle actin. Biochemistry. 1988 Nov 1;27(22):8388–8393. doi: 10.1021/bi00422a014. [DOI] [PubMed] [Google Scholar]
  16. Horiuchi K. Y., Miyata H., Chacko S. Modulation of smooth muscle actomyosin ATPase by thin filament associated proteins. Biochem Biophys Res Commun. 1986 May 14;136(3):962–968. doi: 10.1016/0006-291x(86)90426-2. [DOI] [PubMed] [Google Scholar]
  17. Houk T. W., Jr, Ue K. The measurement of actin concentration in solution: a comparison of methods. Anal Biochem. 1974 Nov;62(1):66–74. doi: 10.1016/0003-2697(74)90367-4. [DOI] [PubMed] [Google Scholar]
  18. Ikebe M., Reardon S. Binding of caldesmon to smooth muscle myosin. J Biol Chem. 1988 Mar 5;263(7):3055–3058. [PubMed] [Google Scholar]
  19. KIELLEY W. W., BRADLEY L. B. The relationship between sulfhydryl groups and the activation of myosin adenosinetriphosphatase. J Biol Chem. 1956 Feb;218(2):653–659. [PubMed] [Google Scholar]
  20. 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]
  21. Klee C. B. Conformational transition accompanying the binding of Ca2+ to the protein activator of 3',5'-cyclic adenosine monophosphate phosphodiesterase. Biochemistry. 1977 Mar 8;16(5):1017–1024. doi: 10.1021/bi00624a033. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Lash J. A., Sellers J. R., Hathaway D. R. The effects of caldesmon on smooth muscle heavy actomeromyosin ATPase activity and binding of heavy meromyosin to actin. J Biol Chem. 1986 Dec 5;261(34):16155–16160. [PubMed] [Google Scholar]
  24. Leavis P. C., Gergely J. Thin filament proteins and thin filament-linked regulation of vertebrate muscle contraction. CRC Crit Rev Biochem. 1984;16(3):235–305. doi: 10.3109/10409238409108717. [DOI] [PubMed] [Google Scholar]
  25. Lehman W. 35 kDa proteins are not components of vertebrate smooth muscle thin filaments. Biochim Biophys Acta. 1989 Jun 13;996(1-2):57–61. doi: 10.1016/0167-4838(89)90094-0. [DOI] [PubMed] [Google Scholar]
  26. Lehman W. Calponin and the composition of smooth muscle thin filaments. J Muscle Res Cell Motil. 1991 Jun;12(3):221–224. doi: 10.1007/BF01745110. [DOI] [PubMed] [Google Scholar]
  27. Lehman W., Sheldon A., Madonia W. Diversity in smooth muscle thin filament composition. Biochim Biophys Acta. 1987 Jul 24;914(1):35–39. doi: 10.1016/0167-4838(87)90158-0. [DOI] [PubMed] [Google Scholar]
  28. Litchfield D. W., Ball E. H. Phosphorylation of caldesmon77 by protein kinase C in vitro and in intact human platelets. J Biol Chem. 1987 Jun 15;262(17):8056–8060. [PubMed] [Google Scholar]
  29. 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]
  30. Marston S. Stoichiometry and stability of caldesmon in native thin filaments from sheep aorta smooth muscle. Biochem J. 1990 Dec 1;272(2):305–310. doi: 10.1042/bj2720305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ngai P. K., Scott-Woo G. C., Lim M. S., Sutherland C., Walsh M. P. Activation of smooth muscle myosin Mg2+-ATPase by native thin filaments and actin/tropomyosin. J Biol Chem. 1987 Apr 15;262(11):5352–5359. [PubMed] [Google Scholar]
  32. 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]
  33. Ngai P. K., Walsh M. P. The effects of phosphorylation of smooth-muscle caldesmon. Biochem J. 1987 Jun 1;244(2):417–425. doi: 10.1042/bj2440417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nishida W., Abe M., Takahashi K., Hiwada K. Do thin filaments of smooth muscle contain calponin? A new method for the preparation. FEBS Lett. 1990 Jul 30;268(1):165–168. doi: 10.1016/0014-5793(90)80999-y. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Pope B. J., Wagner P. D., Weeds A. G. Heterogeneity of myosin heavy chains in subfragment-1 isoenzymes rabbit skeletal myosin. J Mol Biol. 1977 Jan 25;109(3):470–473. doi: 10.1016/s0022-2836(77)80024-7. [DOI] [PubMed] [Google Scholar]
  37. Scott-Woo G. C., Sutherland C., Walsh M. P. Kinase activity associated with caldesmon is Ca2+/calmodulin-dependent kinase II. Biochem J. 1990 Jun 1;268(2):367–370. doi: 10.1042/bj2680367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Smith C. W., Pritchard K., Marston S. B. The mechanism of Ca2+ regulation of vascular smooth muscle thin filaments by caldesmon and calmodulin. J Biol Chem. 1987 Jan 5;262(1):116–122. [PubMed] [Google Scholar]
  39. 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]
  40. Takahashi K., Abe M., Hiwada K., Kokubu T. A novel troponin T-like protein (calponin) in vascular smooth muscle: interaction with tropomyosin paracrystals. J Hypertens Suppl. 1988 Dec;6(4):S40–S43. doi: 10.1097/00004872-198812040-00008. [DOI] [PubMed] [Google Scholar]
  41. Takahashi K., Hiwada K., Kokubu T. Isolation and characterization of a 34,000-dalton calmodulin- and F-actin-binding protein from chicken gizzard smooth muscle. Biochem Biophys Res Commun. 1986 Nov 26;141(1):20–26. doi: 10.1016/s0006-291x(86)80328-x. [DOI] [PubMed] [Google Scholar]
  42. Takahashi K., Hiwada K., Kokubu T. Occurrence of anti-gizzard P34K antibody cross-reactive components in bovine smooth muscles and non-smooth muscle tissues. Life Sci. 1987 Jul 20;41(3):291–296. doi: 10.1016/0024-3205(87)90151-2. [DOI] [PubMed] [Google Scholar]
  43. Takahashi K., Hiwada K., Kokubu T. Vascular smooth muscle calponin. A novel troponin T-like protein. Hypertension. 1988 Jun;11(6 Pt 2):620–626. doi: 10.1161/01.hyp.11.6.620. [DOI] [PubMed] [Google Scholar]
  44. Umekawa H., Hidaka H. Phosphorylation of caldesmon by protein kinase C. Biochem Biophys Res Commun. 1985 Oct 15;132(1):56–62. doi: 10.1016/0006-291x(85)90987-8. [DOI] [PubMed] [Google Scholar]
  45. Vancompernolle K., Gimona M., Herzog M., Van Damme J., Vandekerckhove J., Small V. Isolation and sequence of a tropomyosin-binding fragment of turkey gizzard calponin. FEBS Lett. 1990 Nov 12;274(1-2):146–150. doi: 10.1016/0014-5793(90)81350-w. [DOI] [PubMed] [Google Scholar]
  46. Velaz L., Hemric M. E., Benson C. E., Chalovich J. M. The binding of caldesmon to actin and its effect on the ATPase activity of soluble myosin subfragments in the presence and absence of tropomyosin. J Biol Chem. 1989 Jun 5;264(16):9602–9610. [PubMed] [Google Scholar]
  47. Winder S. J., Walsh M. P. Smooth muscle calponin. Inhibition of actomyosin MgATPase and regulation by phosphorylation. J Biol Chem. 1990 Jun 15;265(17):10148–10155. [PubMed] [Google Scholar]
  48. Woods E. F. Comparative physicochemical studies on vertebrate tropomyosins. Biochemistry. 1969 Nov;8(11):4336–4344. doi: 10.1021/bi00839a017. [DOI] [PubMed] [Google Scholar]
  49. Yamazaki K., Itoh K., Sobue K., Mori T., Shibata N. Purification of caldesmon and myosin light chain (MLC) kinase from arterial smooth muscle: comparisons with gizzard caldesmon and MLC kinase. J Biochem. 1987 Jan;101(1):1–9. doi: 10.1093/oxfordjournals.jbchem.a121879. [DOI] [PubMed] [Google Scholar]
  50. Zot A. S., Potter J. D. Structural aspects of troponin-tropomyosin regulation of skeletal muscle contraction. Annu Rev Biophys Biophys Chem. 1987;16:535–559. doi: 10.1146/annurev.bb.16.060187.002535. [DOI] [PubMed] [Google Scholar]

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