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The kinetic constants for the three substrates were determined for both enzymes using a coupled enzyme assay.
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While the close similarities in both structure and function of FTase and geranylgeranyltransferase type I (GGTase-I) make it likely that both enzymes use a similar catalytic mechanism, it will still be important to conduct detailed studies with GGTase-I to determine whether there are important differences among these enzymes.
Both enzymes use an active site carboxylate to position a water molecule for N-glycosidic bond hydrolysis (E125 in AAG and D238 in AlkA).
Steady-state kinetic studies are carried out on both the wild-type and mutant enzymes using a spectrum of substrates.
The discriminatory power was equal with both enzymes used, which showed a high genetic diversity among C. perfringens.
Both enzymes use inorganic polyphosphate [poly(P)] as a phosphate donor.
Both enzymes use one molecule of hydrogen peroxide to form a high valent iron intermediate named Compound I (Cpd I).
Therefore, both enzymes used in the study were cytosolic proteins.
Digested chromatin was marked by incubating with biotin-14-dCTP and Klenow enzyme using a fill-in reaction20 resulting in blunt-ended repaired DNA strands.
A previously designed enzyme used a reactive lysine to initiate cleavage of a carbon-carbon bond.
Kennedy et al.5,6 first described these enzymes using both an insoluble substrate (diacylglycerol) and a soluble substrate (CDP-alcohol).
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