RESUMO
With yeast-soluble inorganic pyrophosphatase, the heat released during PP(i) hydrolysis was -6.3 kcal/mol regardless of the KCl concentration in the medium. With the membrane-bound pyrophosphatase of corn vacuoles, the heat released varies between -23.5 and -7.5 kcal/mol depending on the KCl concentration in the medium and whether or not a H(+) gradient is formed across the vacuole membranes. The data support the proposal that enzymes are able to handle the energy derived from phosphate compound hydrolysis in such a way as to determine the parcel that is used for work and the fraction that is converted into heat.
Assuntos
Difosfatos/metabolismo , Pirofosfatases/fisiologia , Vacúolos/enzimologia , Leveduras/enzimologia , Zea mays/enzimologia , Catálise , Temperatura Alta , HidróliseRESUMO
Maize root tonoplasts are able to accumulate Ca(2+) using the energy derived from the H(+) gradient formed during PP(i) hydrolysis. Oxalate increases 6- to 10-fold the amount of Ca(2+) accumulated by tonoplast. Two apparently different K(s) values for Ca(2+) with values of 0.36 and 4.70 microM were detected when oxalate was included in the medium and the free Ca(2+) concentration in the medium was buffered with the use of EGTA. Binding of Ca(2+) to the outer surface of tonoplasts inhibits the outflow of Ca(2+) previously accumulated by the tonoplast, half-maximal inhibition being observed in presence of 1 microM Ca(2+). Thapsigargin, a specific inhibitor of Ca(2+)-ATPase, inhibits the Ca(2+) uptake driven by H(+) gradient but does not inhibit the hydrolysis of PP(i) nor the formation of a H(+) gradient.