CHM 4139 Lecture Notes - Lecture 15: Schiff Base, Reaction Intermediate, Enzyme
7 Nov 2016
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Chm 4139- determination of enzyme mechanisms (cont"d: detection of reaction intermediate. An intermediate species (e-x) of an enzyme mechanism (e. g. : an acyl-enzyme intermediate) can be either detected directly (e. g. Covalent modification of the enzyme and each intermediate absorb different wavelength (give different absorbance signal to be measured) *not all has intermediates with different absorbance signal. The intermediate in this case can be isolated by chemically modifying them to make them more stable. During the reaction, a protonated schiff base (imine) is formed. This imine can be stabilized by reduction with nabh4. The resulting amine can be isolated and characterized. Sequence of complex formation during reaction (ordered, random or. Competitive inhibitors can help define the active site. Inhibitors with ki in the picomolar range (10-12 m) are usually structurally similar to the transition state. The tighter the binding, the closer to the transition state structure. Kie is the change in reaction rate due to isotopic substitution.
