... rates of NADPH from wild-type and mutant ecIII determined by fluorescence The release rates of NADPH from ecIII enzymes were determined by fluorescence as described in Materials and Methods The ... purified as described in Materials and methods In addition, the previously made mutants ecIIID392C and ecIIIT393C in loop D, and ecIIIG430C and ecIIIA432C in loop E, were included All mutants were ... (2000) Interactions of the NADP(H)- binding domain III of proton-translocating transhydrogenase from Escherichia coli with NADP (H) and the NAD (H) -binding domain I studied by NMR and site-directed...