In vitro motility assay (IVMA) experiments were performed to analyze the movement of actin filaments sliding on a pavement of myosin molecules at different [ATP] and [ADP]. In standard experimental conditions at [ATP] = 2 mM, about 80% of the actin filaments move in unloaded conditions with a constant velocity. However, a fraction of at least 20% static actin filaments is always present.
View Article and Find Full Text PDFThe thermodynamics of chemical reactions in which all species are explicitly considered with atoms and charge balanced is compared with the transformed thermodynamics generally used to treat biochemical reactions where atoms and charges are not balanced. The transformed thermodynamic quantities suggested by Alberty are obtained by execution of Legendre transformation of the usual thermodynamic potentials. The present analysis demonstrates that the transformed values for ΔG' and ΔH'can be obtained directly without performing Legendre transformations by simply writing the chemical reactions with all the pseudoisomers explicitly included and charges balanced.
View Article and Find Full Text PDFA novel procedure is presented which, by balancing elements and electric charge of biochemical reactions which occur at constant pH and pMg, allows assessing the thermodynamics properties of reaction Δ(r)G'⁰, Δ(r)H'⁰, Δ(r)S'⁰ and the change in binding of hydrogen and magnesium ions of these reactions. This procedure of general applicability avoids the complex calculations required by the use of the Legendre transformed thermodynamic properties of formation Δ(f)G'⁰, Δ(f)H'⁰ and Δ(f)S'⁰ hitherto considered an obligatory prerequisite to deal with the thermodynamics of biochemical reactions. As a consequence, the term "conditional" is proposed in substitution of "Legendre transformed" to indicate these thermodynamics properties.
View Article and Find Full Text PDFThe Legendre-transformed Gibbs energy change for a biochemical reaction, Delta(r)G', is shown to be equal to the nontransformed Gibbs energy change, Delta(r)G, of any single reaction involving selected chemical species of the biochemical system. These two Gibbs energies of reaction have hitherto been thought to have different values. The equality of the quantities means that a substantial part of biochemical and chemical thermodynamics, previously treated separately, can be treated within a unified thermodynamic framework.
View Article and Find Full Text PDFMagnetic Resonance Spectroscopy affords the possibility of assessing in vivo the thermodynamic status of living tissues. The main thermodynamic variables relevant for the knowledge of the health of living tissues are: DeltaG of ATP hydrolysis and cytosolic [ADP], the latter as calculated from the apparent equilibrium constant of the creatine kinase reaction. In this study we assessed the stoichiometric equilibrium constant of the creatine kinase reaction by in vitro (31)P NMR measurements and computer calculations resulting to be: logK(CK)=8.
View Article and Find Full Text PDFThe p K(a) values of 6-fluoro-4,8-diazadodecane-1,12-diamine (6-fluorospermine) (1), 6,6-difluoro-4,8-diazadodecane-1,12-diamine (6,6-difluorospermine) (2), 6-fluoro-4-azaoctane-1,8-diamine (6-fluorospermidine) (3) and 6,6-difluoro-4-azaoctane-1,8-diamine (6,6-difluorospermidine) (4) in D(2)O solution have been determined at 40 degrees C from (13)C NMR chemical shifts data using the new computer program HypNMR2000. The enthalpies of protonation of compounds 1-4 and the parent amines spermine (5) and spermidine (6) have been determined from microcalorimetric titration data. The values of Delta H degrees were used to derive basicity constants relative to 25 degrees C.
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