Inhibition

The rate of reaction can be slowed down or inhibited by competitive and noncompetitive inhibition The two types of inhibitions can be determined by plotting the rate of enzyme activity with the inhibitor and without the presence of inhibitor

Competitive inhibition
The competitive inhibition increases the K. without affecting Vmax Competitive inhibitor and substrate compete for the same active site of the enzyme. A higher substrate concentration is needed to overcome the competitive competition to achieve the same velocity reached in the absence of the competitor. Vmax can be reached if there is sufficient substrate. One half Vmax would require a higher substrate concentration, thus KM is larger.

Non-competitive inhibition
The non-competitive inhibition decreases Vm without affecting i. Non-competitive inhibition cannot be overcome by increasing the substrate concentration. Thus the Vm of the reaction decreases. The non-competitive inhibitor and substrate bind to different sites on the enzyme. The enzyme therefore shows the same KM in the presence or absence of the non-competitive inhibitor

Inhibitor
is a substance that binds to an enzyme, thus, decreasing or stopping its activity. Enzyme inhibitors can be grouped into two types: (a) Competitive inhibitors (b) Non-competitive inhibitors

Competitive inhibitors
increases KM without affecting Vmax Competitive inhibitors have a shape similar to the natural substrate. They can fit temporarily into the active site of the enzyme-preventing substrate binding to it. The competitive inhibitor and natural substrate compete for the same active site of the enzyme. Entry of a competitive inhibitors or substrates would depend on their relative concentration. Competitive inhibition is reversed by increasing the substrate concentration

Examples of competitive inhibitors

Malonate. It competes with succinate for the active site on the enzyme succinate dehydrogenase

Non-competitive inhibitors
decreases Vmi KM is unchanged Non-competitive inhibitors have no structural similarities to the natural substrate. They do not attach to the active site but bind with the enzyme at another site (allosteric site) The binding causes a change in conformation of the enzyme molecule and its active site- prevents the substrate from binding to its active The non-competitive inhibitor and the substrate are not competing for the active site. Therefore an increase in substrate concentration does not affect the rate of reaction

 The rate of reaction decreases with increasing inhibitor concentration. An example of a non-competitive inhibitor is : cyanide which attaches itself to the copper prosthetic group of enzyme cytochrome oxidase and inhibiting respiratory reactions.

End-product inhibition
The end-product of a reaction can act as a non-competitive reversible inhibitor and regulates the metabolic pathway. Example: ATP is produced by cellular respiration.

 When the concentration of ATP is high it acts as an allosteric inhibitor and inhibits biochemical reactions. When ATP concentration falls, ATP leaves the allosteric site and cellular respiration is no longer inhibited

Riversible inhibition

Competitive inhibition is reversible as the inhibitor binds temporarily the active site. It can be overcome by increasing the relativeconcentration of the substrate Some non-competitive inhibitions are reversible, that is, if the inhibitor binds temporarily and loosely to the allosteric site

 Some inhibitors bind very tightly, often, by forming covalent bonds with the enzyme. The nerve gas DIPF is an irreversible inhibitor. It binds permanently with acetylcholinesterase, altering its shape. The enzyme cannot bind with and break down its substrate acetylcholine (neurotransmitter). Acetylcholine molecules accumulate in the synaptic cleft. Nerve impulses cannot be stopped causing continuous muscle contraction. This leads to convulsions, paralysis and eventually, death.

 Many pesticides such as organophosphate pesticides act as irreversible enzyme inhibitors. Exposure to the pesticides can produce harmful effects to the nervous and muscular systems of humans. Heavy metal ions such as Hg2, Ag, As and iodine-containing compounds which combine permanently with sulphydryl groups in the active site or other parts of the enzyme cause inactivation of enzymes.

reversible (rapid binding/release from enzyme in an equilibrium) or irreversible (very tightly bound to enzyme, either covalently or noncovalently, but effectively don't come off) Reversible inhibitors type of inhibition diagnosed by effect of inhibitor on Km and Vmax effects/"diagnosis" obvious on double reciprocal plot. basis for drugs' actions research tools in figuring out enzyme chemical catalytic mechanisms

Reversible Inhibition: competitive uncompetitive, or noncompetitive defined operationally by their effects on enzyme kinetic parameters, Km and/or Vmax.

Reversible Enzyme Inhibitors, continued Decreases Km and reduces Vmax by same factor so slope of 1/Vo vs. 1/[S] doesnt change binds only to ES complex Has no effect on Km (no effect on S binding) reduces Vmax (reduces kcat) Pure B ES Complex Prevents S from binding, so increases Km