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Rationale to measure the forward reaction rate of PGK1. In a reaction mixture containing GA3P, NAD, and Pi, add an excessive amount of GAPDH, bringing the reaction rapidly to equilibrium state, which is monitored at A340. We then added ADP and PGK1 into the reaction mixture to initiate the PGK1-catalyzed forward reaction and recorded the initial rate (see also the text).
