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Metabolism
Suicide substrates (5) are substrate ana-

Inhibitors
Many substances can affect metabolic processes by influencing the activity of enzymes. Enzyme inhibitors are particularly important here. A large proportion of medicines act as enzyme inhibitors. Enzymekinetic experiments are therefore an important aspect of drug development and testing procedures. Natural metabolites are also involved in regulatory processes as inhibitors (see p.114).
A. Types of inhibitor
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Most enzyme inhibitors act reversiblyi.e., they do not cause any permanent changes in the enzyme. However, there are also irreversible inhibitors that permanently modify the target enzyme. The mechanism of action of an inhibitorits inhibition typecan be determined by comparing the kinetics (see p. 92) of the inhibited and uninhibited reactions (B). This makes it possible to distinguish competitive inhibitors (left) from noncompetitive inhibitors (right), for example. Allosteric inhibition is particularly important for metabolic regulation (see below). Substrate analogs (2) have properties similar to those of one of the substrates of the target enzyme. They are bound by the enzyme, but cannot be converted further and therefore reversibly block some of the enzyme molecules present. A higher substrate concentration is therefore needed to achieve a halfmaximum rate; the Michaelis constant Km increases (B). High concentrations of the substrate displace the inhibitor again. The maximum rate Vmax is therefore not influenced by this type of inhibition. Because the substrate and the inhibitor compete with one another for the same binding site on the enzyme, this type of inhibition is referred to as competitive. Analogs of the transition state (3) usually also act competitively. When an inhibitor interacts with a group that is important for enzyme activity, but does not affect binding of the substrate, the inhibition is non-competitive (right). In this case, Km remains unchanged, but the concentration of functional enzyme [E]t, and thus Vmax, decrease. Non-competitive inhibitors generally act irreversibly, by modifying functional groups of the target enzyme (4).

logs that also contain a reactive group. Initially, they bind reversibly, and then they form a covalent bond with the active center of the enzyme. Their effect is therefore also non-competitive. A well-known example of this is the antibiotic penicillin (see p. 254). Allosteric inhibitors bind to a separate binding site outside the active center ( 6). This results in a conformational change in the enzyme protein that indirectly reduces its activity (see p.116). Allosteric effects practically only occur in oligomeric enzymes. The kinetics of this type of system can no longer be described using the simple Micha elisMenten model.
B. Inhibition kinetics
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In addition to the LineweaverBurk plot (see p.92), the EadieHofstee plot is also commonly used. In this case, the velocity v is plotted against v /[A]. In this type of plot, Vmax corresponds to the intersection of the approximation lines with the v axis, while Km is derived from the gradient of the lines. Competitive and non-competitive inhibitors are also easily distinguishable in the Eadie Hofstee plot. As mentioned earlier, competitive inhibitors only influence K m, and not Vmax. The lines obtained in the absence and presence of an inhibitor therefore intersect on the ordinate. Non-competitive inhibitors produce lines that have the same slope (Km unchanged) but intersect with the ordinate at a lower level. Another type of inhibitor, not shown here, in which Vmax and Km are reduced by the same factor, is referred to as uncompetitive. Inhibitors with purely uncompetitive effects are rare. A possible explanation for this type of inhibition is selective binding of the inhibitor to the EA complex. Allosteric enzymes shift the target enzymes saturation curve to the left (see p. 92). In EadieHofstee and LineweaverBurk plots (see p. 92), allosteric enzymes are recognizable because they produce curved lines (not shown).

Koolman, Color Atlas of Biochemistry, 2nd edition 2005 Thieme All rights reserved. Usage subject to terms and conditions of license.