Allostery
 
Allostery is used to describe regulatory phenomena in biological systems. The term “allosteric” (other shape) was coined by Monod and coworkers to describe a particular type of regulatory behavior and marks an important synthesis in the development of biochemistry and molecular biology. Since its inception, the term has evolved to describe several related concepts and is used today to describe a variety of phenomena. To some, it is associated with a particular type of regulatory behavior, while to many it is used to describe several different aspects of regulation. The development of the concept, the meanings it has come to have, its application in describing regulatory properties of proteins and enzymes, and recent developments that indicate the need to use it in its original sense are described here.
The allosteric concept was conceived in the early 1960s by Monod and coworkers to describe the properties of regulatory enzymes. In the mid-1950s, several investigators described enzymes whose catalytic properties were dependent on effector molecules other than the substrate. The first well-characterized example of this behavior was the finding by Cori and colleagues that 5′-AMP is required for the in vitro catalytic activity of glycogen phosphorylase b from resting skeletal muscle (1). Studies of the regulation of biosynthetic pathways in bacteria showed that the catalytic properties of the first enzyme in a pathway are often modulated by the end product of the pathway, which has a structure different from that of the substrate—for example, isoleucine inhibition of threonine deaminase (see Threonine Operon) (2) and CTP inhibition of aspartate transcarbamoylase (3). The differences in the structures of the substrates and effectors were first noted by Monod and Jacob, who postulated in a seminal paper in 1961 that the effectors bind to separate and distinct sites (4). They coined the term allosteric site for the effector binding site to distinguish it from the active site and postulated that the effectors act indirectly by causing changes in the conformation of the enzyme that alter its catalytic site and kinetic properties. In 1963, Monod and coworkers compiled the available data on regulatory enzymes (5). These enzymes all contained more than one subunit per enzyme molecule–that is, dimers, tetramers, and dodecamers. For several of these enzymes, changes in quaternary structure (association–dissociation of the subunits) had been shown to occur upon addition of the effectors. Monod was also aware of unpublished crystallographic results showing that the oxygen-binding sites on hemoglobin are located far from one another and that the distances between the amino acid residues labeled by heavy atoms change upon binding of oxygen (Ref. 6, pp. 577(–578. On this basis, the key elements of a general model for the functional structures of regulatory enzymes were formulated:
1. Each molecule of the native enzyme contains multiple subunits with binding sites for substrates and ligands, and the regulatory behavior depends on the relationships between the subunits.
2. No direct interactions between substrate(s) or effectors are required, that is, the effectors act indirectly on the catalytic site.
3. The actions of the effectors are due entirely to a reversible conformational change in the protein that is induced by effector binding.
Monod et al. postulated that protein conformational change is the basis for control and coordination of chemical events in living cells, and Monod considered this to be the “second secret of life” (Ref. 6, p. 576). Structural studies using X-ray crystallography methods support the concepts of this general model.
These concepts were used to develop the concerted model, a specific molecular model for describing regulatory behavior (7). This model was developed as a plausible explanation for the positive cooperativity in the binding of oxygen to hemoglobins. Because the apparent affinity for binding of oxygen changes as its concentration is varied, these are called homotropic interactions. The concerted model retains the aspects of the general model with respect to oligomeric structure of the protein, multiple binding sites located far from one another, and conformational changes upon binding of one ligand that are transmitted by protein conformational changes to the other ligand binding sites, thereby altering the binding affinities. This model defines the relations between the conformational changes and ligand binding. The word concerted refers to the all-or-none nature of the conformational transition. That is, only two conformations are allowed for the subunits in an oligomer and all of the subunits in a given oligomer have the same conformation. The conformations are denoted by the terms R-state and T-state, and they differ in affinity for binding ligands. The R-state has the higher affinity for ligands. Positive cooperativity in oxygen binding is explained by a large predominance of the unliganded protein in the T-state with a shift to the R-state upon binding of oxygen. The model was extended to enzymes by assuming that the interactions between the substrate and enzyme are in rapid equilibrium and that all forms of the enzyme have the same Vmax. At this juncture, the term allosteric takes a different meaning. The conformational change between the two states is called an allosteric transition. So, in addition to referring to sites for molecules other than the substrate, it refers to interactions between substrate binding sites on different subunits. The concept of the “other shape” is extended to the enzymes, which are termed “allosteric enzymes.”
This initial formulation of the concerted model describes homotropic interactions. The original considerations of the allosteric concept were developed to treat cases in which the behavior of one molecule, the substrate, depends on the concentration of another molecule. These are called heterotropic interactions. The regulatory enzymes whose properties were catalogued by Monod et al. showed both homotropic and heterotropic effects (5). The phenomenon of desensitization—that is, treatments that result in loss of substrate homotropic interactions and heterotropic interactions, but do not affect catalytic activity, had been described for some of these enzymes. Monod et al. postulated that desensitization means that the molecular basis might be the same for both homotropic and heterotropic interactions. They stated that if this were the case, the concerted model could also be used to describe heterotropic effects—that is, the original allostery. In the model, activating effectors have higher affinity for the R-state, while inhibitory effectors have higher affinity for the T-state. This issue has resulted in great confusion in the use of the model. All subsequent treatments presume that a single allosteric transition is the basis for both homotropic and heterotropic effects. This presumption is particularly evident in structural studies of regulatory proteins, where the terms R-state and T-state dominate discussions.
In the three cases of regulatory proteins that have been examined with care, it is clear that the concerted model fails. More than two states are involved in oxygen binding to hemoglobin (8, 9). Homotropic and heterotropic interactions are not due to the same transition in aspartate transcarbamoylase (10, 11). Although the same T-state is predicted for phosphofructokinase with different inhibitors, experimental results show that the properties of the inhibited forms are different (12). These results argue strongly that the term allosteric should be used in a stricter sense to refer to effects of one ligand on the binding of another, independently of assumptions about the mechanism of the interaction.
References
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6.H. F. Judson (1979) The Eighth Day of Creation: The Makers of the Revolution in Biology, Simon & Shuster, New York. 
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11. R. C. Stevens, Y. M. Chook, C. Y. W. N. Cho, Lipscomb, and E. R. Kantrowitz (1991) Protein. Eng. 4, 391–408. 
12. V. L. Tlapak-Simmons and G. D. Reinhart (1994) Arch. Biochem. Biophys. 308, 226–230.