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. 1989 Apr;55(4):595–603. doi: 10.1016/S0006-3495(89)82857-7

Role of MgATP and MgADP in the cross-bridge kinetics in chemically skinned rabbit psoas fibers. Study of a fast exponential process (C)

M Kawai 1, H R Halvorson 1
PMCID: PMC1330542  PMID: 2785822

Abstract

The role of the substrate (MgATP) and product (MgADP) molecules in cross-bridge kinetics is investigated by small amplitude length oscillations (peak to peak: 3 nm/cross-bridge) and by following amplitude change and phase shift in tension time courses. The range of discrete frequencies used for this investigation is 0.25-250 Hz, which corresponds to 0.6-600 ms in time domain. This report investigates the identity of the high frequency exponential advance (process C), which is equivalent to "phase 2" of step analysis. The experiments are performed in maximally activated (pCa 4.5-5.0) single fibers from chemically skinned rabbit psoas fibers at 20 degrees C and at the ionic strength 195 mM. The rate constant 2 pi c deduced from process (C) increases and saturates hyperbolically with an increase in MgATP concentration, whereas the same rate constant decreases monotonically with an increase in MgADP concentration. The effects of MgATP and MgADP are opposite in all respects we have studied. These observations are consistent with a cross-bridge scheme in which MgATP and MgADP are in rapid equilibria with rigorlike cross-bridges, and they compete for the substrate site on myosin heads. From our measurements, the association constants are found to be 1.4 mM-1 for MgATP and 2.8 mM-1 for MgADP. We further deduced that the composite second order rate constant of MgATP binding to cross-bridges and subsequent isomerization/dissociation reaction to be 0.57 x 10(6)M-1s-1.

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Selected References

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  1. Arata T., Mukohata Y., Tonomura Y. Structure and function of the two heads of the myosin molecule. VI. ATP hydrolysis, shortening, and tension development of myofibrils. J Biochem. 1977 Sep;82(3):801–812. doi: 10.1093/oxfordjournals.jbchem.a131756. [DOI] [PubMed] [Google Scholar]
  2. Brenner B., Yu L. C., Greene L. E., Eisenberg E., Schoenberg M. Ca2+-sensitive cross-bridge dissociation in the presence of magnesium pyrophosphate in skinned rabbit psoas fibers. Biophys J. 1986 Dec;50(6):1101–1108. doi: 10.1016/S0006-3495(86)83554-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cooke R., Crowder M. S., Thomas D. D. Orientation of spin labels attached to cross-bridges in contracting muscle fibres. Nature. 1982 Dec 23;300(5894):776–778. doi: 10.1038/300776a0. [DOI] [PubMed] [Google Scholar]
  4. Cooke R., Pate E. The effects of ADP and phosphate on the contraction of muscle fibers. Biophys J. 1985 Nov;48(5):789–798. doi: 10.1016/S0006-3495(85)83837-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Curtin N. A., Gilbert C., Kretzschmar K. M., Wilkie D. R. The effect of the performance of work on total energy output and metabolism during muscular contraction. J Physiol. 1974 May;238(3):455–472. doi: 10.1113/jphysiol.1974.sp010537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feit H., Kawai M., Schulman M. I. Stiffness and contractile properties of avian normal and dystrophic muscle bundles as measured by sinusoidal length perturbations. Muscle Nerve. 1985 Jul-Aug;8(6):503–510. doi: 10.1002/mus.880080605. [DOI] [PubMed] [Google Scholar]
  7. Geeves M. A., Jeffries T. E., Millar N. C. ATP-induced dissociation of rabbit skeletal actomyosin subfragment 1. Characterization of an isomerization of the ternary acto-S1-ATP complex. Biochemistry. 1986 Dec 30;25(26):8454–8458. doi: 10.1021/bi00374a020. [DOI] [PubMed] [Google Scholar]
  8. Glyn H., Sleep J. Dependence of adenosine triphosphatase activity of rabbit psoas muscle fibres and myofibrils on substrate concentration. J Physiol. 1985 Aug;365:259–276. doi: 10.1113/jphysiol.1985.sp015770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goldman Y. E., Hibberd M. G., McCray J. A., Trentham D. R. Relaxation of muscle fibres by photolysis of caged ATP. Nature. 1982 Dec 23;300(5894):701–705. doi: 10.1038/300701a0. [DOI] [PubMed] [Google Scholar]
  10. Goldman Y. E., Hibberd M. G., Trentham D. R. Initiation of active contraction by photogeneration of adenosine-5'-triphosphate in rabbit psoas muscle fibres. J Physiol. 1984 Sep;354:605–624. doi: 10.1113/jphysiol.1984.sp015395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldman Y. E. Kinetics of the actomyosin ATPase in muscle fibers. Annu Rev Physiol. 1987;49:637–654. doi: 10.1146/annurev.ph.49.030187.003225. [DOI] [PubMed] [Google Scholar]
  12. Greene L. E., Eisenberg E. Cooperative binding of myosin subfragment-1 to the actin-troponin-tropomyosin complex. Proc Natl Acad Sci U S A. 1980 May;77(5):2616–2620. doi: 10.1073/pnas.77.5.2616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Greene L. E., Eisenberg E. Dissociation of the actin.subfragment 1 complex by adenyl-5'-yl imidodiphosphate, ADP, and PPi. J Biol Chem. 1980 Jan 25;255(2):543–548. [PubMed] [Google Scholar]
  14. Greene L. E., Sellers J. R., Eisenberg E., Adelstein R. S. Binding of gizzard smooth muscle myosin subfragment 1 to actin in the presence and absence of adenosine 5'-triphosphate. Biochemistry. 1983 Feb 1;22(3):530–535. doi: 10.1021/bi00272a002. [DOI] [PubMed] [Google Scholar]
  15. Hammes G. G. Relaxation spectrometry of biological systems. Adv Protein Chem. 1968;23:1–57. doi: 10.1016/s0065-3233(08)60399-x. [DOI] [PubMed] [Google Scholar]
  16. Heinl P., Kuhn H. J., Rüegg J. C. Tension responses to quick length changes of glycerinated skeletal muscle fibres from the frog and tortoise. J Physiol. 1974 Mar;237(2):243–258. doi: 10.1113/jphysiol.1974.sp010480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Highsmith S. Interactions of the actin and nucleotide binding sites on myosin subfragment 1. J Biol Chem. 1976 Oct 25;251(20):6170–6172. [PubMed] [Google Scholar]
  18. Huxley A. F., Simmons R. M. Proposed mechanism of force generation in striated muscle. Nature. 1971 Oct 22;233(5321):533–538. doi: 10.1038/233533a0. [DOI] [PubMed] [Google Scholar]
  19. Johnson K. A., Taylor E. W. Intermediate states of subfragment 1 and actosubfragment 1 ATPase: reevaluation of the mechanism. Biochemistry. 1978 Aug 22;17(17):3432–3442. doi: 10.1021/bi00610a002. [DOI] [PubMed] [Google Scholar]
  20. Johnson R. E., Adams P. H. ADP binds similarly to rigor muscle myofibrils and to actomyosin-subfragment one. FEBS Lett. 1984 Aug 20;174(1):11–14. doi: 10.1016/0014-5793(84)81067-4. [DOI] [PubMed] [Google Scholar]
  21. Kawai M., Brandt P. W. Sinusoidal analysis: a high resolution method for correlating biochemical reactions with physiological processes in activated skeletal muscles of rabbit, frog and crayfish. J Muscle Res Cell Motil. 1980 Sep;1(3):279–303. doi: 10.1007/BF00711932. [DOI] [PubMed] [Google Scholar]
  22. Kawai M., Güth K., Winnikes K., Haist C., Rüegg J. C. The effect of inorganic phosphate on the ATP hydrolysis rate and the tension transients in chemically skinned rabbit psoas fibers. Pflugers Arch. 1987 Jan;408(1):1–9. doi: 10.1007/BF00581833. [DOI] [PubMed] [Google Scholar]
  23. Kawai M. Head rotation or dissociation? A study of exponential rate processes in chemically skinned rabbit muscle fibers when MgATP concentration is changed. Biophys J. 1978 Apr;22(1):97–103. doi: 10.1016/S0006-3495(78)85473-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kawai M., Schachat F. H. Differences in the transient response of fast and slow skeletal muscle fibers. Correlations between complex modulus and myosin light chains. Biophys J. 1984 Jun;45(6):1145–1151. doi: 10.1016/S0006-3495(84)84262-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kawai M. The role of orthophosphate in crossbridge kinetics in chemically skinned rabbit psoas fibres as detected with sinusoidal and step length alterations. J Muscle Res Cell Motil. 1986 Oct;7(5):421–434. doi: 10.1007/BF01753585. [DOI] [PubMed] [Google Scholar]
  26. Levy R. M., Umazume Y., Kushmerick M. J. Ca2+ dependence of tension and ADP production in segments of chemically skinned muscle fibers. Biochim Biophys Acta. 1976 May 14;430(2):352–365. doi: 10.1016/0005-2728(76)90091-8. [DOI] [PubMed] [Google Scholar]
  27. Lienhard G. E., Secemski I. I. P 1 ,P 5 -Di(adenosine-5')pentaphosphate, a potent multisubstrate inhibitor of adenylate kinase. J Biol Chem. 1973 Feb 10;248(3):1121–1123. [PubMed] [Google Scholar]
  28. Lymn R. W., Taylor E. W. Mechanism of adenosine triphosphate hydrolysis by actomyosin. Biochemistry. 1971 Dec 7;10(25):4617–4624. doi: 10.1021/bi00801a004. [DOI] [PubMed] [Google Scholar]
  29. Marston S. B., Taylor E. W. Comparison of the myosin and actomyosin ATPase mechanisms of the four types of vertebrate muscles. J Mol Biol. 1980 Jun 5;139(4):573–600. doi: 10.1016/0022-2836(80)90050-9. [DOI] [PubMed] [Google Scholar]
  30. Millar N. C., Geeves M. A. Protein fluorescence changes associated with ATP and adenosine 5'-[gamma-thio]triphosphate binding to skeletal muscle myosin subfragment 1 and actomyosin subfragment 1. Biochem J. 1988 Feb 1;249(3):735–743. doi: 10.1042/bj2490735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Millar N. C., Geeves M. A. The limiting rate of the ATP-mediated dissociation of actin from rabbit skeletal muscle myosin subfragment 1. FEBS Lett. 1983 Aug 22;160(1-2):141–148. doi: 10.1016/0014-5793(83)80954-5. [DOI] [PubMed] [Google Scholar]
  32. Pate E., Cooke R. Energetics of the actomyosin bond in the filament array of muscle fibers. Biophys J. 1988 Apr;53(4):561–573. doi: 10.1016/S0006-3495(88)83136-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schoenberg M. Characterization of the myosin adenosine triphosphate (M.ATP) crossbridge in rabbit and frog skeletal muscle fibers. Biophys J. 1988 Jul;54(1):135–148. doi: 10.1016/S0006-3495(88)82938-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schoenberg M., Eisenberg E. ADP binding to myosin cross-bridges and its effect on the cross-bridge detachment rate constants. J Gen Physiol. 1987 Jun;89(6):905–920. doi: 10.1085/jgp.89.6.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sleep J. A., Hutton R. L. Exchange between inorganic phosphate and adenosine 5'-triphosphate in the medium by actomyosin subfragment 1. Biochemistry. 1980 Apr 1;19(7):1276–1283. doi: 10.1021/bi00548a002. [DOI] [PubMed] [Google Scholar]
  36. Sugi H., Kobayashi T. Sarcomere length and tension changes in tetanized frog muscle fibers after quick stretches and releases. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6422–6425. doi: 10.1073/pnas.80.20.6422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Takashi R., Putnam S. A fluorimetric method for continuously assaying ATPase: application to small specimens of glycerol-extracted muscle fibers. Anal Biochem. 1979 Jan 15;92(2):375–382. doi: 10.1016/0003-2697(79)90674-2. [DOI] [PubMed] [Google Scholar]
  38. Thomas D. D. Spectroscopic probes of muscle cross-bridge rotation. Annu Rev Physiol. 1987;49:691–709. doi: 10.1146/annurev.ph.49.030187.003355. [DOI] [PubMed] [Google Scholar]
  39. Trybus K. M., Taylor E. W. Transient kinetics of adenosine 5'-diphosphate and adenosine 5'-(beta, gamma-imidotriphosphate) binding to subfragment 1 and actosubfragment 1. Biochemistry. 1982 Mar 16;21(6):1284–1294. doi: 10.1021/bi00535a028. [DOI] [PubMed] [Google Scholar]
  40. White H. D., Taylor E. W. Energetics and mechanism of actomyosin adenosine triphosphatase. Biochemistry. 1976 Dec 28;15(26):5818–5826. doi: 10.1021/bi00671a020. [DOI] [PubMed] [Google Scholar]
  41. Yanagida T. Angles of nucleotides bound to cross-bridges in glycerinated muscle fiber at various concentrations of epsilon-ATP, epsilon-ADP and epsilon-AMPPNP detected by polarized fluorescence. J Mol Biol. 1981 Mar 15;146(4):539–560. doi: 10.1016/0022-2836(81)90046-2. [DOI] [PubMed] [Google Scholar]

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