Influence of inorganic phosphate and pH on sarcoplasmic reticular ATPase in skinned muscle fibres of Xenopus laevis.
1. The influence of 30 mM inorganic phosphate (Pi) and pH (6.2-7.4) on the rate of ATP utilization was determined in mechanically skinned bundles of myofibrils from the iliofibularis muscle of Xenopus laevis at approximately 5 C. 2. BDM (2,3-butanedione monoxime; 10 mM) depressed isometric force production and actomyosin (AM) ATPase activity equally. Therefore sarcoplasmic reticular (SR) ATPase activity could be determined by extrapolation of the total ATPase activity to zero force. 3. The SR ATPase activity without added Pi at pH 7.1 was 42 +/- 2 % of the total ATPase activity. Addition of 30 mM Pi reduced SR ATPase activity slightly, by 9 +/- 5 %, and depressed force by 62 +/- 2 % and AM ATPase activity by 21 +/- 6 %. 4. At pH 6.2, force, SR ATPase activity and AM ATPase activity were reduced by 21 +/- 5, 61 +/- 5 and 10 +/- 4 % of their respective values at pH 7.1. 5. The SR ATPase activity at 30 mM Pi and pH 6.2 was reduced markedly to 20 +/- 6 % of the value under control conditions, suggesting that the maximum rate of Ca2+ uptake during muscle fatigue was strongly depressed. This reduction was larger than expected on the basis of the effects of Pi and pH alone.
Laboratory for Physiology, Institute for Cardiovascular Research, Free University, van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands. gjm.stienen.physiol@med.vu.nl
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GJ
Stienen
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Papp
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Zaremba
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D003931:Diacetyl / Q000031:analogs & derivatives / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006863:Hydrogen-Ion Concentration; D007537:Isometric Contraction / Q000187:drug effects; D007700:Kinetics; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D009210:Myofibrils / Q000187:drug effects / Q000201:enzymology; D010710:Phosphates / Q000494:pharmacology*; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D014982:Xenopus laevis
D004791:Enzyme Inhibitors; D010710:Phosphates; C004717:diacetylmonoxime; D000251:Adenosine Triphosphatases; D003931:Diacetyl