Effects of temperature, ionic strength and pH on the function of skeletal muscle myosin from a eurythermal fish, Fundulus heteroclitus.
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 microm s(-1) at 26 degrees C, ionic strength (Gamma/2) of 0.08 M Gamma/2 and pH 7.4. Speed increased as temperature increased over the range of 5-36 degrees C with an activation energy (E (a)) of 94.0+/-7.0 kJ mol(-1)) and an enthalpy (DeltaH (double dagger)) of 91.5+/-7.0 kJ mol(-1) at 20 degrees C. A linear relationship between temperature and ATPase activity was also obtained with actin-activated myosin Mg(2+)-ATPase assays over the temperature range 5-35 degrees C with E (a=)59.9+/-2.4 kJ mol(-1) and DeltaH (double dagger)=57.4+/-2.4 kJ mol(-1) at 20 degrees C. There was little or no effect of ionic strength on filament speed over the range 0.19 M Gamma/2-0.54 M Gamma/2. Speed increased significantly at lower ionic strengths and was 7.9-fold higher at 0.08 M Gamma/2 than at 0.19 M Gamma/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.
Grove
Theresa J
TJ
Department of Biological Science, Biology Unit 1, Florida State University, Tallahassee, Florida 32306-4370, USA. grove@bio.fsu.edu
McFadden
Lori A
LA
Chase
P Bryant
PB
Moerland
Timothy S
TS
D008841:Actin Cytoskeleton / Q000737:chemistry / Q000502:physiology; D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D023781:Fundulidae / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D009119:Muscle Contraction / Q000502:physiology; D018482:Muscle, Skeletal / Q000737:chemistry*; D009218:Myosins / Q000737:chemistry / Q000502:physiology*; D009994:Osmolar Concentration; D011817:Rabbits; D013696:Temperature
D000199:Actins; D009218:Myosins