The following data was collected for an unknown reaction mechanism for an enzyme

Question

The following data was collected for an unknown reaction mechanism for an enzyme catalyzing two substrates. The second substrate concentrates were as follows. 1.00 x 10^-3 M for experiment 1, 2.00 x 10^-3 M for experiment 2, 3.00 x 10^-3 M for experiment 3. Determine whether the reaction is a sequential or ping-pong mechanism, and determine of the kinetic constants associated with that type of reaction mechanism

1 IS1] v1 (M/s) v2 (M/s) v3 (M/s) 0.002 0.0203251165 0.0403222815 0.0602403324 0.004 0.0203250297 0.0403219823 0.0602397010 0.006 0.0203249430 0.0403216832 0.0602390696 0.008 0.0203248562 0.0403213840 0.0602384382 0.01 0.0203247695 0.0403210849 0.0602378068 0.012 0.0203246827 0.0403207857 0.060237175 10 12 13

Explanation / Answer

We have sevaral reaction rates of the enzymatic reaction with varying concentration of Substrate-1 keeping the value of substrate- 2 constant. We also have the rates of the reaction measured similarly with different constant concentration of substrate-2. We can easily find out what the mechanism of the enzymatic reaction is- ping pong mechanism or sequebtial mechanism by Lineweaver-Burk/double reciprocal plot.

Plot 1/V vs. 1/S-1, with a different line for each concentration of S-2 . If the lines representing different constant S-2 concentration intersects, then it is sequential mechanism. If the lines are parallel, it indicates that the reactio follows ping-pong mechanism.

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