Abstract
The mummichog, Fundulus heteroclitus, is an intertidal fish that exhibits little change in swimming ability despite large and rapid variations in environmental parameters. We therefore tested the hypothesis that this nearly constant function is due to Fundulus myosin being intrinsically insensitive to changes of temperature, ionic strength and pH. In vitro motility assays were used to quantify the speed of unregulated actin filaments on myosin purified from F. heteroclitus glycolytic skeletal muscle. Filament speed was 2.07±0.17 μm s−1 at 26°C, ionic strength (Γ/2) of 0.08 M Γ/2 and pH 7.4. Speed increased as temperature increased over the range of 5–36°C with an activation energy (E a) of 94.0±7.0 kJ mol−1) and an enthalpy (ΔH ‡) of 91.5±7.0 kJ mol−1 at 20°C. A linear relationship between temperature and ATPase activity was also obtained with actin-activated myosin Mg2+-ATPase assays over the temperature range 5–35°C with E a=59.9±2.4 kJ mol−1 and ΔH ‡=57.4±2.4 kJ mol−1 at 20°C. There was little or no effect of ionic strength on filament speed over the range 0.19 M Γ/2–0.54 M Γ/2. Speed increased significantly at lower ionic strengths and was 7.9-fold higher at 0.08 M Γ/2 than at 0.19 M Γ/2. Speed increased with pH with a 16-fold increase between pH 6.7 and 7.4. These results indicate that changes in physiological parameters that include temperature, pH and ionic strength affect the function of unregulated F. heteroclitus myosin, and thus other factors must be responsible for the mummichog’s swimming performance being comparatively insensitive to environmental variation.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Anson M, (1992) Temperature dependence and Arrhenius activation energy of F-actin velocity generated in vitro by skeletal myosin J Mol Biol 224: 1029–1038
Armitage P, (1980) Statistical Methods in Medical Research. Blackwell Scientific Research. Blackwell Scientific Publications Oxford, UK p. 279–301
Chaen S, Nakaya M, Guo X-F, Watabe S, (1996) Lower activation energy for sliding of F-actin on a less thermostable isoform of carp myosin J Biochem 120: 788–791
Chase PB, Kushmerick MJ, (1988) Effects of pH on contraction of rabbit fast and slow skeletal muscle fibers Biophys J 53: 935–946
Chase PB, Chen Y, Kulin K, Daniel TL, (2000) Viscosity and solute dependence of F-actin translocation by rabbit skeletal heavy meromyosin Am J Physiol 278: C1088–C1098l
Chidester FE, (1920) The behavior of Fundulus heteroclitus on Salt Marshes of New Jersey Am Naturalist 54: 551–557
Gordon AM, LaMadrid MA, Chen Y, Luo Z, Chase PB, (1997) Calcium regulation of skeletal muscle thin filament motility in vitro Biophys J 72: 1295–1307
Hartshorne DJ, Barns EM, Parker L, Fuchs F, (1972) The effect of temperature on actomyosin Biochim Biophys Acta 267: 190–202
Homsher E, Wang F, Sellers JR, (1992) Factors affecting movement of F-actin filaments propelled by skeletal muscle heavy meromyosin Am J Physiol 262: C714–C723
Homsher E, Wang F, Sellers JR, (1993) Factors affecting filament velocity in in vitro motility assays and their relation to unloaded shortening velocity in muscle fibers, In: Sug H, Pollach GH, (eds.) Mechanism of Myofilament Sliding in Muscle Contraction Plenum Press New York. (pp. 279–290)
Homsher E, Kim B, Bobkova A, Tobacman LS, (1996) Calcium regulation of thin filament movement in an in vitro motility assay Biophys J 70: 1881–1892
Hwang GC, Watabe S, Hashimoto K, (1990) Changes in carp myosin ATPase induced by temperature acclimation J Comp Physiol B 160: 233–239
Johnson T, Bennett A, (1995) The thermal acclimation of burst escape performance in fish: an integrated study of molecular and cellular physiology and organismal performance J Exp Biol 198: 2165–2175
Johnston IA, Goldspink G, (1975) Thermodynamic activation parameters of fish myofibrillar ATPase enzyme and evolutionary adaptations to temperature Nature 257: 620–622
Johnston IA, Walesby NJ, (1977) Molecular mechanisms of temperature adaptation in fish myofibrillar adenosine triphosphatases J Comp Physiol 119: 195–206
Jones RH, Molitoris BA, (1984) A statistical method for determining the breakpoint of two lines Anal Biochem 141: 287–290
Julian FJ, Moss RL, (1981) Effects of calcium and ionic strength on shortening velocity and tension development in frog skinned muscle fibers J Physiol 311: 179–199
Kron SJ, Toyoshima YY, Uyeda TQ, Spudich JA, (1991) Assays for actin sliding movement over myosin-coated surfaces Meth Enzymol 196: 399–416
Margossian SS, Lowey S, (1982) Preparation of myosin and its subfragments from rabbit skeletal muscle Meth Enzymol 85B: 55–71
Moerland TS, Egginton S, (1998) Intracellular pH of muscle and temperature: insight from in vivo 31P NMR measurements in a stenothermal Antarctic teleost (Harpagifer antarcticus) J Therm Biol 23: 275–282
Pardee JD, Spudich JA, (1982) Purification of muscle actin Meth Enzymol 85B: 164–181
Peyser YM, Ajtai K, Burghardt TP, Muhlrad A, (2001) Effect of ionic strength on the conformation of myosin subfragment 1-nucleotide complexes Biophys J 81: 1101–1114
Prosser CL (1973) Inorganic ions. In: Prosser CL (ed.) Comparative Animal Physiology, 3rd edn. (pp. 79–110). W.B. Saunders Company, Philadelphia
Reeves RB, (1972) An imidazole alphastat hypothesis for vertebrate acid-base regulation: tissue carbon dioxide content and body temperature in bullfrogs Resp Physiol 14: 219–236
Sartoris FJ, Bock C, Pörtner HO, (2003) Temperature-dependent pH regulation in eurythermal and stenothermal marine fish: an interspecies comparison using 31P-NMR J Thermal Biol 28: 3632–371
Schägger H, von Jagow G, (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa Anal Biochem 166: 368–379
Seow CY, Ford LE, (1993) High ionic strength and low pH detain activated skinned rabbit skeletal muscle crossbridges in a low force state J Gen Physiol 101: 487–511
Sidell BD, Johnston IA, Moerland TS, Goldspink G, (1983) The eurythermal myofibrillar protein complex of the mummichog: adaptation to a functional thermal environment J Comp Physiol 153: 167–173
Siemankowski RF, Wiseman MO, White HD, (1985) ADP dissociation from actomyosin subfragment 1 is sufficiently slow to limit the unloaded shortening velocity in vertebrate muscle Proc Natl Acad Sci USA 82: 658–662
Stein LA, Chock PB, Eisenberg E, (1984) The rate-limiting step in the actomyosin adenosine triphosphatase cycle Biochemistry 23: 1555–1563
von Hippel PH, Wong KY, (1964) Neutral Salts: the generality of their effects on the stability of macromolecular conformations Science 145: 577–580
White HD, (1982) Special instrumentation and techniques for kinetic studies of contractile systems Meth Enzymol 85B: 698–708
Willmer P, Stone G, Johnston I, (2000) Environmental Physiology of Animals Blackwell Science, London (pp. 57–84)
Acknowledgements
We are extremely grateful to Shanedah Williams Brown for assistance with actin purification. We thank Thomas Asbury for writing the Linux based motion analysis program.
This work is sponsored by NSF grant number IBN-98-08120 and by the Defense Advanced Research Projects Agency (DARPA) Defense Sciences Office (DSO) under the auspices of Drs Alan Rudolph and Anantha Krishnan through the Space and Naval Warfare Systems Center (SPAWAR), San Diego, contract number N66001-02-C-8030.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Grove, T.J., McFadden, L.A., Chase, P.B. et al. Effects of temperature, ionic strength and pH on the function of skeletal muscle myosin from a eurythermal fish, Fundulus heteroclitus . J Muscle Res Cell Motil 26, 191–197 (2005). https://doi.org/10.1007/s10974-005-9010-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10974-005-9010-0