Quantitative mathematical expressions for accurate in vivo assessment of cytosolic [ADP] and DeltaG of ATP hydrolysis in the human brain and skeletal muscle |
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Authors: | Iotti Stefano Frassineti Chiara Sabatini Antonio Vacca Alberto Barbiroli Bruno |
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Institution: | Biochimica Clinica, Dipartimento di Medicina Clinica e Biotecnologia Applicata "D. Campanacci", Università di Bologna, Via Massarenti, 9, 40138 Bologna, Italy. stefano.iotti@unibo.it |
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Abstract: | Magnetic 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)=8.00+/-0.07 at T=310 K and ionic strength I=0.25 M. This value refers to the equilibrium: PCr(2-)+ADP(3-)+ H(+)=Cr+ATP(4-). We also assessed by computer calculation the stoichiometric equilibrium constant of ATP hydrolysis obtaining the value: logK(ATP-hyd)=-12.45 at T=310 K and ionic strength I=0.25 M, which refers to the equilibrium: ATP(4-)+H(2)O=ADP(3-)+PO(4)(3-)+2H(+). Finally, we formulated novel quantitative mathematical expressions of DeltaG of ATP hydrolysis and of the apparent equilibrium constant of the creatine kinase reaction as a function of total PCr], pH and pMg, all quantities measurable by in vivo (31)P MRS. Our novel mathematical expressions allow the in vivo assessment of cytosolic ADP] and DeltaG of ATP hydrolysis in the human brain and skeletal muscle taking into account pH and pMg changes occurring in living tissues both in physiological and pathological conditions. |
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