Biopolym. Cell. 1995; 11(5):28-36.
Caldesmon is a Ca2+-regulatory protein component of native aorta smooth muscle filaments
1Danilova V. M., 1Kulikova N. V., 1Tregubov V. S., 1Omelyanuk V. S., 1Filenko A. M.
  1. Petr Bogach Institute of Physiology
    Taras Shevchenko National University of Kyiv
    2, Academika Glushkova Ave Str., Kyiv, Ukraine, 03187

Abstract

The techniques of preparation and purification of calmodulin- and actin-binding protein caldesmon (CaD) from pig aorta smooth muscle is described. It is established that CaD prevents the interaction of myosin and actin thus inhibiting the Mg2+-ATPase activity of actomyosin irrespectively of Ca2+ concentration. The inhibitory effect of CaD is cancelled at addition of calmodulin in the presence of Ca2+. With the help of spectro-fluorimetry it is shown that calmodulin may bind to caldesmon causing conformational changes in its structure. Both the results obtained and the data known from literature suggest CaD to be a likely regulator of vascular smooth muscle actomyosin system.

References

[1] Marston SB. The regulation of smooth muscle contractile proteins. Progr Biophys Mol Biol. 1983;41:1–41.
[2] Marston SB, Taylor EW. Comparison of the myosin and actomyosin ATPase mechanisms of the four types of vertebrate muscles. J Mol Biol. 1980;139(4):573-600.
[3] Ebashi S. The Croonian lecture, 1979: Regulation of muscle contraction. Proc R Soc Lond B Biol Sci. 1980;207(1168):259-86.
[4] Lehman W, Szent-Györgyi AG. Regulation of muscular contraction. Distribution of actin control and myosin control in the animal kingdom. J Gen Physiol. 1975;66(1):1-30.
[5] Marston SB, Trevett RM, Walters M. Calcium ion-regulated thin filaments from vascular smooth muscle. Biochem J. 1980;185(2):355-65.
[6] Danilova VM. Regulation of actin-myosin interaction in smooth muscles of vertebrates. Mechanisms of control of muscle activity. L. Nauka, 1985: 128-47.
[7] Walsh MP. The Ayerst Award Lecture 1990. Calcium-dependent mechanisms of regulation of smooth muscle contraction. Biochem Cell Biol. 1991;69(12):771-800.
[8] Khokhlova VS, Kulikova NV, Tregubov VS, Danilova VM. The role of myosin phosphorylation in the regulation of smooth muscle actin-myosin interaction. Mol Gen Biophys. 1991; 16:74-78.
[9] Marston SB, Smith CW. The thin filaments of smooth muscles. J Muscle Res Cell Motil. 1985;6(6):669-708.
[10] Lehman W. Calponin and the composition of smooth muscle thin filaments. J Muscle Res Cell Motil. 1991;12(3):221-4.
[11] 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;78(9):5652-5.
[12] Bryan J. Caldesmon, acidic amino acids and molecular weight determinations. J Muscle Res Cell Motil. 1989;10(2):95-6.
[13] 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;101(1):1-9.
[14] Marston SB, Redwood CS. The molecular anatomy of caldesmon. Biochem J. 1991;279 ( Pt 1):1-16.
[15] Szpacenko A, Dabrowska R. Functional domain of caldesmon. FEBS Lett. 1986;202(2):182-6.
[16] Fujii T, Imai M, Rosenfeld GC, Bryan J. Domain mapping of chicken gizzard caldesmon. J Biol Chem. 1987;262(6):2757-63.
[17] Bartegi A, Fattoum A, Derancourt J, Kassab R. Characterization of the carboxyl-terminal 10-kDa cyanogen bromide fragment of caldesmon as an actin-calmodulin-binding region. J Biol Chem. 1990;265(25):15231-8. PubMed
[18] Katayama E, Horiuchi KY, Chacko S. Characteristics of the myosin and tropomyosin binding regions of the smooth muscle caldesmon. Biochem Biophys Res Commun. 1989;160(3):1316-22.
[19] Makuch R, Walsh MP, Dabrowska R. Location of the calmodulin- and actin-binding domains at the C-terminus of caldesmon. FEBS Lett. 1989;247(2):411–4.
[20] Mabuchi K, Wang CL. Electron microscopic studies of chicken gizzard caldesmon and its complex with calmodulin. J Muscle Res Cell Motil. 1991;12(2):145-51.
[21] 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;842(1):70-5.
[22] Bretscher A. Smooth muscle caldesmon. Rapid purification and F-actin cross-linking properties. J Biol Chem. 1984;259(20):12873-80.
[23] Khokhlova VS, Tregubov VS, Danilova VM. Two types of Ca2+ -dependent regulation of actin-myosin interaction in smooth muscles of the stomach of a pig. Mol Gen Biophys. 1986; 11: 28-33.
[24] Khaitlina SY. Methods for isolation and purification of actin. Assessment of purity and nativity. Biophysics. and biochem. methods of muscle proteins. L. Nauka. 1978: 122-141.
[25] Danilova VM, Tregubov VS Comparative study of structural and functional properties of myosin of smooth and skeletal muscle of mammals. Mol Gen Biophys. 1988; 13:12-21.
[26] Teo TS, Wang TH, Wang JH. Purification and properties of the protein activator of bovine heart cyclic adenosine 3',5'-monophosphate phosphodiesterase. J Biol Chem. 1973;248(2):588-95.
[27] Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.
[28] Tregubov VS, Sopin GE. Universal electrophoretic system for the separation of proteins with high resolution. Mol Gen Biophys. 1988. 13: 125-129.
[29] Marston SB, Lehman W. Caldesmon is a Ca2+-regulatory component of native smooth-muscle thin filaments. Biochem J. 1985;231(3):517-22.
[30] Marston SB, Redwood CS. The essential role of tropomyosin in cooperative regulation of smooth muscle thin filament activity by caldesmon. J Biol Chem. 1993;268(17):12317-20.
[31] Sobue K, Kanda K, Tanaka T, Ueki N. Caldesmon: a common actin-linked regulatory protein in the smooth muscle and nonmuscle contractile system. J Cell Biochem. 1988;37(3):317-25.
[32] missed
[33] Levine BA, Moir AJ, Audemard E, Mornet D, Patchell VB, Perry SV. Structural study of gizzard caldesmon and its interaction with actin. Binding involves residues of actin also recognised by myosin subfragment 1. Eur J Biochem. 1990;193(3):687-96.
[34] Bartegi A, Fattoum A, Kassab R. Cross-linking of smooth muscle caldesmon to the NH2-terminal region of skeletal F-actin. J Biol Chem. 1990;265(4):2231-7.
[35] Adams S, DasGupta G, Chalovich JM, Reisler E. Immunochemical evidence for the binding of caldesmon to the NH2-terminal segment of actin. J Biol Chem. 1990;265(32):19652-7.
[36] Nowak E, Borovikov YS, Dabrowska R. Caldesmon weakens the bonding between myosin heads and actin in ghost fibers. Biochim Biophys Acta. 1989;999(3):289-92.
[37] Chalovich JM, Yu LC, Brenner B. Involvement of weak binding crossbridges in force production in muscle. J Muscle Res Cell Motil. 1991;12(6):503-6.
[38] Marston S. Aorta caldesmon inhibits actin activation of thiophosphorylated heavy meromyosin Mg2+-ATPase activity by slowing the rate of product release. FEBS Lett. 1988;238(1):147-50.
[39] Marston SB, Redwood CS. Inhibition of actin-tropomyosin activation of myosin MgATPase activity by the smooth muscle regulatory protein caldesmon. J Biol Chem. 1992;267(24):16796-800.