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The effect of Ca2+ on the structure of synthetic filaments of smooth muscle myosin

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Abstract

Using electron microscopy and negative staining we have studied the effect of Ca2+ on the structure of synthetic filaments of chicken gizzard smooth muscle myosin under conditions applied by Frado and Craig (1989) for demonstration of the influence of Ca2+ on the structure of synthetic filaments of scallop striated muscle myosin. The results show that Ca2+ induces the transition of compact, ordered structure of filaments with a 14.5 nm axial repeat of the myosin heads close to the filament backbone (characteristic of the relaxing conditions) to a disordered structure with randomly arranged myosin heads together with subfragments-2 (S-2) seen at a distance of up to 50 nm from the filament backbone. This order/disorder transition is much more pronounced in filaments formed of unphosphorylated myosin, since a substantial fraction of phosphorylated filaments in the relaxing solution is already disordered due to phosphorylation. Under rigor conditions some of the filaments of unphosphorylated and phosphorylated myosin retain a certain degree of order resembling those under relaxing conditions, while most of them have a substantially disordered appearance. The results indicate that Ca2+-induced movement of myosin heads away from the filament backbone is an inherent property of smooth muscle myosin, like molluscan muscle myosin regulated exclusively by Ca2+ binding, and can play a modulatory role in smooth muscle contraction.

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References

  • Allen BG and Walsh MP (1994) The biochemical basis of the regulation of smooth muscle contraction. Trends BiochemSci 19: 362–368.

    Google Scholar 

  • Barsotti RJ, Ikebe Mand, Hartshorne DJ (1987) Effects of Ca2., Mg2. and myosin phosphorylation on skinned smooth muscle fibers. Am J Physiol 252: C543–554.

    Google Scholar 

  • Chalovich JM and Pfitzer G (1997) Structure and function of the thin filament proteins of smooth muscle. In: Kao CY and Carsten ME (teds). Cellular Aspects of Smooth Muscle Function (pp.253–286). Cambridge University Press, Cambridge, UK.

    Google Scholar 

  • Clarke ML, Hofmann W and Wray JS (1986) ATP binding and crossbridge structure in muscle. J Mol Biol 191: 581–585.

    Google Scholar 

  • Cooke PH, Fay FS and Craig R (1989) Myosin filaments isolated from skinned amphibian smooth muscle cells are side–polar. J Muscle Res Cell Motil 10: 206–220.

    Google Scholar 

  • Craig R, Padron R and Kendrick–Jones J (1987) Structural changes accompanying phosphorylation of tarantula muscle myosin filaments. J Cell Biol 184: 429–443.

    Google Scholar 

  • Da Silva ACR and Reinach FC (1991) Calcium binding induces conformational changes in muscle regulatory proteins. Trends Biochem Sci 16: 53–57.

    Google Scholar 

  • Da Silva ACR, Kendrick–Jones J and Reinach FC (1995) Determinants of ion specificity on EF–hands sites. J Biol Chem 270: 67736778.

    Google Scholar 

  • Dąbrowska R (1994) Actin and thin filament–associated proteins in smooth muscle. In: Raeburn D and Giembycz MA (eds). Airways Smooth Muscle: Biochemical Control of Contraction and Relaxation (pp. 31–59). Birkhäuser Verlag, Basel.

    Google Scholar 

  • Diffee GM, Greaser ML, Reinach FC and Moss RL (1995) Effects of non–divalent cation binding mutant of myosin regulatory light chain on tension generation in skinned skeletal muscle fibers. Biophys J 68: 1443–1452.

    Google Scholar 

  • Diffee GM, Patel JR, Reinach FC, Greaser ML and Moss RL (1996) Altered kinetics of contraction in skeletal muscle fibers containing a mutant myosin regulatory light chain with reduced divalent cation binding. Biophys J 71: 341–350.

    Google Scholar 

  • Ebashi S (1991) Excitation–contraction coupling and the mechanism of muscle contraction. Annu Rev Physiol 53: 1–16.

    Google Scholar 

  • Farah CS and Reinach FC (1995) The troponin complex and regulation of muscle contraction. FASEB J 9: 755–776.

    Google Scholar 

  • Frado L–LY and Craig RJ (1989) Structural changes induced in Ca2.regulated myosin filaments by Ca2. and ATP. J Cell Biol 109: 529–538.

    Google Scholar 

  • Hai CM and Murphy RA (1989) Ca2., crossbridge phosphorylation, and contraction. Annu Rev Physiol 51: 285–298.

    Google Scholar 

  • Hartshorne DJ and Kawamura T (1992) The biochemical basis of the regulation of smooth muscle contraction. News Physiol Sci 7: 59–63.

    Google Scholar 

  • Heaslip RJ and Chacko S (1985) Effects of Ca2. and Mg2. on the actomyosin adenosine–50–triphosphatase of stably phosphorylated gizzard myosin. Biochemistry 24: 2731–2736.

    Google Scholar 

  • Holroyde MJ, Potter JD and Solaro RJ (1979) The calcium binding properties of phosphorylated and unphosphorylated cardiac and skeletal myosins. J Biol Chem 254: 6478–6482.

    Google Scholar 

  • Ikebe M and Hartshorne DJ (1985)Effects of Ca2. on the conformation and enzymatic activity of smooth muscle myosin. J Biol Chem 260: 13146–13153.

    Google Scholar 

  • Ikebe M and Ogihara S (1982) Phosphorylation–dependent and ATPinduced changes in structural array in gizzard myosin filament bundles. J Biochem (Tokyo) 92: 1973–1977.

    Google Scholar 

  • Jakes R, Northrop F and Kendrick–Jones J (1976) Calcium binding regions of myosin regulatory light chains. FEBS Lett 70: 229–232.

    Google Scholar 

  • Kamm KE and Stull JT (1985) The function of myosin and myosin light chain kinase phosphorylation in smooth muscle. Annu Rev Pharmacol Toxicol 25: 593–620.

    Google Scholar 

  • Knight PJ (1996) Dynamic behavior of the head–tail junction of myosin. J Mol Biol 255: 269–274.

    Google Scholar 

  • Kulikova N and Dąbrowska R (1996) The influence of caldesmon on papain proteolysis of monomeric smooth muscle myosin. Biochem Biophys Res Commun 225: 195–202.

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Google Scholar 

  • Levine RJC (1997) Differences in myosin head arrangement on relaxed thick filaments from Lethocerus and rabbit muscles. J Muscle Res Cell Motil 18: 529–543.

    Google Scholar 

  • Levine RJC, Kensler RW, Yang Z, Stull JT and Sweeney HL (1996) Myosin light chain phosphorylation affects the structure of rabbit skeletal muscle thick filaments. Biophys J 71: 898–907.

    Google Scholar 

  • Levine RJC, Yang Z, Epstein ND, Fananapazir L, Stull JT and Sweeney HL (1998) Structural and functional responses of mammalian thick filaments to alterations in myosin regulatory light chains. J Struct Biol 122: 149–161.

    Google Scholar 

  • Lowey S and Risby D (1971) Light chains from fast and slow muscle myosin. Nature 234: 81–85.

    Google Scholar 

  • Marston SB and Redwood CS (1991) The molecular anatomy of caldesmon. Biochem J 279: 1–16.

    Google Scholar 

  • Méenéetret J–P, Hofmann W and Lepault J (1988) Cryo–electron microscopy of insect flight muscle thick filaments. An approach to dynamic electron microscope studies. J Mol Biol 202: 175–178.

    Google Scholar 

  • Morris EP, Squire JM and Fuller GW (1991) The 4–stranded arrangement of myosin heads on insects (Lethocerus) flight muscle thick filaments. J Struct Biol 107: 237–249.

    Google Scholar 

  • Moss RL (1992) Ca2. regulation of mechanical properties of striated muscle. Mechanistic studies using extraction and replacement of regulatory proteins. Circ Res 70: 865–884.

    Google Scholar 

  • Notarianni G, Collins J, Leszyk J and Marston SB (1998) A novel Ca2.–binding protein associated with caldesmon in smooth muscle thin filaments. J Muscle Res Cell Motil 19: 311.

    Google Scholar 

  • Podlubnaya Z, Stępkowski D, Udaltsov SN and Kąkol I (1994) Attachment–detachment behavior of crossbridges on the surface of reconstituted myosin filaments: effect of pH, Ca–ions and myosin light chains phosphorylation. J Muscle Res Cell Motil 15: 203.

    Google Scholar 

  • Podlubnaya Z, Kąkol I, Moczarska A and Stępkowski D (1996) Structural behavior of crossbridges in reconstituted rabbit cardiac myosin filaments: effect of pH–value and Ca2.–level. J Muscle Res Cell Motil 17: 117.

    Google Scholar 

  • Podlubnaya Z, Kulikova N and Dąbrowska R (1999) Ca2.–binding to smooth muscle myosin changes the structure and enzymatic properties of its reconstituted filaments. J Muscle Res Cell Motil. 20: 81–82.

    Google Scholar 

  • Rayment I, Rypniewski WR, Schmidt–Bäse K, Smith R, Tomchick DR, Benning MM, Winkelmann DA, Wesenberg G and Holden HM (1993) Three–dimensional structure of myosin subfragment–1: a molecular motor. Science 261: 50–58.

    Google Scholar 

  • Rees DD and Frederiksen DW (1981) Calcium regulation of porcine aortic myosin. J Biol Chem 256: 357–364.

    Google Scholar 

  • Siegman MJ, Butler TM, Mooers SU and Michaøek A (1984) Ca2. can affect Vmax without changes in myosin light chain phosphorylation in smooth muscle. PfluÈgers Arch 401: 385–390.

    Google Scholar 

  • Sobieszek A (1994) Smooth muscle myosin: molecule conformation, filament assembly and associated regulatory enzymes. In: Raeburn D and Giembycz MA (eds). Airways Smooth Muscle: Biochemical Control of Contraction and Relaxation (pp. 1–29). Birkhäuser Verlag, Basel.

    Google Scholar 

  • Sobieszek A (1985) Phosphorylation reaction of vertebrate smooth muscle myosin: an enzyme kinetic analysis. Biochemistry 24: 12661274.

    Google Scholar 

  • Sobieszek A and Jertschin P (1986) Urea–glycerol–acrylamide gel electrophoresis of acidic low molecular weight muscle proteins: rapid determination of myosin light chain phosphorylation in myosin, actomyosin and whole muscle samples. Electrophoresis 7: 417–425.

    Google Scholar 

  • Somlyo AP and Somlyo AV (1994) Signal transduction and regulation in smooth muscle. Nature 372: 231–236.

    Google Scholar 

  • Squire JM and Morris EP (1998) A new look at thin filament regulation in vertebrate skeletal muscle. FASEB J 12: 761–771.

    Google Scholar 

  • Stępkowski D, Efimova N, Paczyńska A, Moczarska A, Nieznańska H and Kąkol I (1997) The possible role of myosin A1 light chain in the weakening of actin–myosin interaction. Biochim Biophys Acta 1340: 105–114.

    Google Scholar 

  • Strynadka NCJ and James MNG (1989) Crystal structures of the helix–loop–helix calcium–binding proteins. Annu Rev Biochem 58: 951–998.

    Google Scholar 

  • Swartz DR, Moss RL and Greaser ML (1996) Calcium alone does not fully activate the thin filament for S1 binding to rigor myofibrils. Biophys J 71: 1891–1904.

    Google Scholar 

  • Sweeney HL, Browman BF and Stull JT (1993) Myosin light chain phosphorylation in vertebrate striated muscle: regulation and function. Am J Physiol 264: C1085–C1095.

    Google Scholar 

  • Szczęsna D, Zhao J and Potter JD (1996) The regulatory light chains of myosin modulate cross–bridge cycling in skeletal muscle. J Biol Chem 271: 5246–5250.

    Google Scholar 

  • Szczęsna D, Zhao J, Roman N, Zhi G, Stull J and Potter J (1997) Phosphorylation and Ca2. binding to the regulatory light chains of myosin modulate force development in skeletal muscle. Conference `A Myosin Family Reunion: Biology to Pathology', Bethesda, MD Abstract F–26.

  • Trybus KM (1994) Role of myosin light chains. J Muscle Res Cell Motil 15: 587–594.

    Google Scholar 

  • Trybus KM, Freyzon Y, Faust LZ and Sweeney HL (1997) Spare the rod, spoil the regulation: necessity for a myosin rod. Proc Natl Acad Sci USA 94: 48–52.

    Google Scholar 

  • Trybus KM and Lowey S (1988) The regulatory light chain is required for folding of smooth muscle myosin. J Biol Chem 263: 1648516492.

    Google Scholar 

  • Vibert P and Craig R (1985) Structural changes that occur in scallop myosin filaments upon activation. J Cell Biol 101: 830–837.

    Google Scholar 

  • Watterson JG, Kohler L and Schaub M (1979) Evidence for two distinct anities in the binding of divalent metal ions to myosin. J Biol Chem 254: 6470–6477.

    Google Scholar 

  • Xie X, Harrison DH, Schlichting I, Sweet RM, Kalabokis VN, SzentGyórgyi AG and Cohen C (1994) Structure of the regulatory domain of scallop myosin at 2.8 A Ê resolution. Nature 368: 306–312.

    Google Scholar 

  • Xu J–Q, Harder BA, Uman P and Craig R (1996) Myosin filament structure in vertebrate smooth muscle. J Cell Biol 134: 53–66.

    Google Scholar 

  • Yang Z, Stull JT, Levine RJC and Sweeney HL (1998)Changes in interfilament spacing mimic the effects of myosin regulatory light chain phosphorylation in rabbit psoas fibers. J Struct Biol 122: 139–148. 554

    Google Scholar 

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Authors and Affiliations

  1. Institute of Experimental and Theoretical Biophysics, Russian Academy of Sciences, Pushchino, Russia

    Zoya Podlubnaya

  2. Nencki Institute of Experimental Biology, Polish Academy of Sciences, 3 Pasteur Street, 02-093, Warsaw, Poland

    Natalia Kulikova & Renata Dąbrowska

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  1. Zoya Podlubnaya
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  2. Natalia Kulikova
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  3. Renata Dąbrowska
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Podlubnaya, Z., Kulikova, N. & Dąbrowska, R. The effect of Ca2+ on the structure of synthetic filaments of smooth muscle myosin. J Muscle Res Cell Motil 20, 547–554 (1999). https://doi.org/10.1023/A:1005533020784

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