CHM 4139 Lecture Notes - Lecture 13: Enzyme, Table Tennis, Isomerase
7 Nov 2016
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Document Summary
Coupled assays typically show a lag phase at the beginning. This is the time needed for the coupling enzyme reaction to reach the steady state. *to use coupled assays, 2 conditions must be met: the coupling enzyme reaction cannot be the rate-limiting step. Normally we aim to have around 100 times more of the 2nd enzyme than the 1st: the measured signal should be proportional to the amount of product of the 1st reaction. The most commonly used coupling enzymes are oxidoreductases, in particular those that use the nad(h)/nadp(h) coenzyme. Aka: dehydrogenase due to the ease of its detection. A useful way to represent enzyme reactions is by using a cleland diagram. Tells us what is being bound in what order and what is being releases in which order. And the vertical arrows represents compounds being bound or released by the enzyme during the reaction. To describe these multi-substrate reactions, we use the cleland nomenclature.
