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• W2040610147 abstract "Millisecond mixing and quenching experiments were performed in order to study the rate of phosphorylation by Pi of the Ca2+-dependent ATPase of sarcoplasmic reticulum vesicles. A rapid phosphoenzyme formation was observed when the vesicles were preincubated in the absence of Ca2+ prior to the addition of Pi and Mg2+ to the medium, the half-time being in the range of 6 to 10 ms. A lag phase and a 5- to 10-fold slower rate of phosphoenzyme formation were observed when the enzyme was preincubated with Ca2+ prior to the addition to the reaction mixture of Pi, Mg2+, and an excess of ethylene glycol bis(β-aminoethyl ether)N,N′-tetraacetic acid. The rate of phosphoenzyme hydrolysis was measured either by the addition of Ca2+ or, in the absence of Ca2+, by tracing the hydrolysis of radioactive phosphoenzyme upon the addition of nonradioactive Pi. In the presence of Ca2+, the rate of phosphoenzyme hydrolysis was found to be one order of magnitude slower than the rate of hydrolysis measured in the absence of Ca2+. Different rates of phosphoenzyme formation and cleavage were found depending on whether sarcoplasmic reticulum vesicles or purified Ca2+-dependent ATPase were used. A transient phosphorylation by Pi was observed when the enzyme was preincubated in the absence of Ca2+ and then added to a medium containing Pi, Mg2+, and excess of Ca2+. The enzyme was phosphorylated during the initial 100 ms, the phosphoenzyme formed being slowly hydrolyzed in the subsequent incubation intervals. In these conditions ATP synthesis was observed if ADP was added to the mixture 100 ms after starting the reaction. No transient phosphorylation by Pi was observed when the enzyme was preincubated with Ca2+. Synthesis of a small but significant amount of ATP was observed when the enzyme was preincubated in the absence of Ca2+ and then added to a medium containing Pi, ADP, Mg2+, and 20 mm CaCl2. This was not observed when the enzyme was preincubated in the presence of Ca2+." @default.
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• W2040610147 date "1980-08-01" @default.
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• W2040610147 title "Pathway for ATP synthesis by sarcoplasmic reticulum ATPase" @default.
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• W2040610147 doi "https://doi.org/10.1016/0003-9861(80)90192-7" @default.
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