SUMMARY

10.1 Aspartate Transcarbamoylase Is Allosterically Inhibited by the End Product of Its Pathway

Allosteric proteins constitute an important class of proteins whose biological activity can be regulated. Specific regulatory molecules can modulate the activity of allosteric proteins by binding to distinct regulatory sites, separate from the functional sites. These proteins have multiple functional sites, which display cooperativity as evidenced by a sigmoidal dependence of function on substrate concentration. Aspartate transcarbamoylase (ATCase), one of the best-understood allosteric enzymes, catalyzes the synthesis of N-carbamoylaspartate, the first intermediate in the synthesis of pyrimidines. ATCase is feedback inhibited by CTP, the final product of the pathway. ATP reverses this inhibition. ATCase consists of separable catalytic (c3) subunits (which bind the substrates) and regulatory (r2) subunits (which bind CTP and ATP). The inhibitory effect of CTP, the stimulatory action of ATP, and the cooperative binding of substrates are mediated by large changes in quaternary structure. On binding substrates, the c3 subunits of the c6r6 enzyme move apart and reorient themselves. This allosteric transition is highly concerted. All subunits of an ATCase molecule simultaneously interconvert from the T (low-affinity) to the R (high-affinity) state.

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10.2 Isozymes Provide a Means of Regulation Specific to Distinct Tissues and Developmental Stages

Isozymes differ in structural characteristics but catalyze the same reaction. They provide a means of fine-tuning metabolism to meet the needs of a given tissue or developmental stage. The results of gene-duplication events provide the means for subtle regulation of enzyme function.

10.3 Covalent Modification Is a Means of Regulating Enzyme Activity

The covalent modification of proteins is a potent means of controlling the activity of enzymes and other proteins. Phosphorylation is a common type of reversible covalent modification. Signals can be highly amplified by phosphorylation because a single kinase can act on many target molecules. The regulatory actions of protein kinases are reversed by protein phosphatases, which catalyze the hydrolysis of attached phosphoryl groups.

Cyclic AMP serves as an intracellular messenger in the transduction of many hormonal and sensory stimuli. Cyclic AMP switches on protein kinase A, a major multifunctional kinase, by binding to the regulatory subunit of the enzyme, thereby releasing the active catalytic subunits of PKA. In the absence of cAMP, the catalytic sites of PKA are occupied by pseudosubstrate sequences of the regulatory subunit.

10.4 Many Enzymes Are Activated by Specific Proteolytic Cleavage

The activation of an enzyme by the proteolytic cleavage of one or a few peptide bonds is a recurring control mechanism seen in processes as diverse as the activation of digestive enzymes and blood clotting. The inactive precursor is a zymogen (proenzyme). Trypsinogen is activated by enteropeptidase or trypsin, and trypsin then activates a host of other zymogens, leading to the digestion of foodstuffs. For instance, trypsin converts chymotrypsinogen, a zymogen, into active chymotrypsin by hydrolyzing a single peptide bond.

A striking feature of the clotting process is that it is accomplished by a cascade of zymogen conversions, in which the activated form of one clotting factor catalyzes the activation of the next precursor. Many of the activated clotting factors are serine proteases. In the final step of clot formation, fibrinogen, a highly soluble molecule in the plasma, is converted by thrombin into fibrin by the hydrolysis of four arginine–glycine bonds. The resulting fibrin monomer spontaneously forms long, insoluble fibers called fibrin. Zymogen activation is also essential in the lysis of clots. Plasminogen is converted into plasmin, a serine protease that cleaves fibrin, by tissue-type plasminogen activator. Although zymogen activation is irreversible, specific inhibitors of some proteases exert control. The irreversible protein inhibitor antithrombin III holds blood clotting in check in the clotting cascade.

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