Catalytic inhibitors

Extending Agent, inhibitors. Catalytic Solution and Special Additives)... 

Action patterns Adhesion domains Affinity and mechanism-based inhibitors Catalytic domains Classification schemes Hydrolases Lyases Mechanism of action Phosphorylases Proteinaceous inhibitors... 

Inhibitor, catalytic A species that decreases the rate of an enzyme-catalyzed reaction. 

Catalyst Temperature, Inhibitor Catalytic activity (turnovers/hour)... 

The catalytic subunit of cAPK contains two domains connected by a peptide linker. ATP binds in a deep cleft between the two domains. Presently, crystal structures showed cAPK in three different conformations, (1) in a closed conformation in the ternary complex with ATP or other tight-binding ligands and a peptide inhibitor PKI(5-24), (2) in an intermediate conformation in the binary complex with adenosine, and (3) in an open conformation in the binary complex of mammalian cAPK with PKI(5-24). Fig.l shows a superposition of the three protein kinase configurations to visualize the type of conformational movement. 

The procedure is computationally efficient. For example, for the catalytic subunit of the mammalian cAMP-dependent protein kinase and its inhibitor, with 370 residues and 131 titratable groups, an entire calculation requires 10 hours on an SGI 02 workstation with a 175 MHz MIPS RIOOOO processor. The bulk of the computer time is spent on the FDPB calculations. The speed of the procedure is important, because it makes it possible to collect results on many systems and with many different sets of parameters in a reasonable amount of time. Thus, improvements to the method can be made based on a broad sampling of systems. 

The catalytic subunit then catalyzes the direct transfer of the 7-phosphate of ATP (visible as small beads at the end of ATP) to its peptide substrate. Catalysis takes place in the cleft between the two domains. Mutual orientation and position of these two lobes can be classified as either closed or open, for a review of the structures and function see e.g. [36]. The presented structure shows a closed conformation. Both the apoenzyme and the binary complex of the porcine C-subunit with di-iodinated inhibitor peptide represent the crystal structure in an open conformation [37] resulting from an overall rotation of the small lobe relative to the large lobe. 

Karlsson, R., Zheng, J., Zheng, N.-H., Taylor, S. S., Sowadski, J. M. Structure of the mamalian catalytic subunit of cAMP-dependent protein kinase and an inhibitor peptide displays an open conformation. Acta Cryst. D 49 (1993) 381-388. 

Elucidating Mechanisms for the Inhibition of Enzyme Catalysis An inhibitor interacts with an enzyme in a manner that decreases the enzyme s catalytic efficiency. Examples of inhibitors include some drugs and poisons. Irreversible inhibitors covalently bind to the enzyme s active site, producing a permanent loss in catalytic efficiency even when the inhibitor s concentration is decreased. Reversible inhibitors form noncovalent complexes with the enzyme, thereby causing a temporary de-... 

Some of the physical properties of fatty acid nitriles are Hsted in Table 14 (see also Carboxylic acids). Eatty acid nitriles are produced as intermediates for a large variety of amines and amides. Estimated U.S. production capacity (1980) was >140, 000 t/yr. Eatty acid nitriles are produced from the corresponding acids by a catalytic reaction with ammonia in the Hquid phase. They have Httie use other than as intermediates but could have some utility as surfactants (qv), mst inhibitors, and plastici2ers (qv). 

When the operating temperature exceeds ca 93°C, the catalytic effects of metals become an important factor in promoting oil oxidation. Inhibitors that reduce this catalytic effect usually react with the surfaces of the metals to form protective coatings (see Metal surface treatments). Typical metal deactivators are the zinc dithiophosphates which also decompose hydroperoxides at temperatures above 93°C. Other metal deactivators include triazole and thiodiazole derivatives. Some copper salts intentionally put into lubricants counteract or reduce the catalytic effect of metals. 

Complexing agents, which act as buffers to help control the pH and maintain control over the free metal—salt ions available to the solution and hence the ion concentration, include citric acid, sodium citrate, and sodium acetate potassium tartrate ammonium chloride. Stabilizers, which act as catalytic inhibitors that retard the spontaneous decomposition of the bath, include fluoride compounds thiourea, sodium cyanide, and urea. Stabilizers are typically not present in amounts exceeding 10 ppm. The pH of the bath is adjusted. 

Additive inhibitors have been developed to reduce the contaminant coke produced through nickel-cataly2ed reactions. These inhibitors are injected into the feed stream going to the catalytic cracker. The additive forms a nickel complex that deposits the nickel on the catalyst in a less catalyticaHy active state. The first such additive was an antimony compound developed and first used in 1976 by Phillips Petroleum. The use of the antimony additive reportedly reduced coke yields by 15% in a commercial trial (17). 

Addition Chlorination. Chlorination of olefins such as ethylene, by the addition of chlorine, is a commercially important process and can be carried out either as a catalytic vapor- or Hquid-phase process (16). The reaction is influenced by light, the walls of the reactor vessel, and inhibitors such as oxygen, and proceeds by a radical-chain mechanism. Ionic addition mechanisms can be maximized and accelerated by the use of a Lewis acid such as ferric chloride, aluminum chloride, antimony pentachloride, or cupric chloride. A typical commercial process for the preparation of 1,2-dichloroethane is the chlorination of ethylene at 40—50°C in the presence of ferric chloride (17). The introduction of 5% air to the chlorine feed prevents unwanted substitution chlorination of the 1,2-dichloroethane to generate by-product l,l,2-trichloroethane. The addition of chlorine to tetrachloroethylene using photochemical conditions has been investigated (18). This chlorination, which is strongly inhibited by oxygen, probably proceeds by a radical-chain mechanism as shown in equations 9—13. 

Chlorination of Methane. Methane can be chlorinated thermally, photochemicaHy, or catalyticaHy. Thermal chlorination, the most difficult method, may be carried out in the absence of light or catalysts. It is a free-radical chain reaction limited by the presence of oxygen and other free-radical inhibitors. The first step in the reaction is the thermal dissociation of the chlorine molecules for which the activation energy is about 84 kj/mol (20 kcal/mol), which is 33 kJ (8 kcal) higher than for catalytic chlorination. This dissociation occurs sufficiendy rapidly in the 400 to 500°C temperature range. The chlorine atoms react with methane to form hydrogen chloride and a methyl radical. The methyl radical in turn reacts with a chlorine molecule to form methyl chloride and another chlorine atom that can continue the reaction. The methane raw material may be natural gas, coke oven gas, or gas from petroleum refining. 

Fig. 1. Inhibition of porcine pancreatic a-amylase. Substrates, an inhibitor, and their binding orientations in the active site are shown schematically. The arrows denote the catalytic site in each case, (a) The small substrate, G2PNP [17400-77-0] (3) (b) the large substrate, G OH [13532-61 -1] (4) and (c) the inhibitor, 4-phenyl imidazole (5) and the substrate G2PNP (3) in the binding orientation for noncompetitive inhibition. The binding orientation of G2PNP...

The efficiency of inactivation by covalent bond formation vs release of the reactive species into solution has been described by its partition ratio. The most efficient inactivators have catalytic partition ratios of 0, in which case each inhibitor molecule leads to inactivation of the enzyme. To this date, many of these inhibitors have been designed, and alternative names like suicide substrate, Trojan Horse inactivator, enzyme induced inactivator, inhibitor, and latent inactivator have been used for this class of inhibitors. A number of comprehensive reviews are available (26—32). 

Usually, a rapid binding step of the inhibitor I to the enzyme E leads to the formation of the initial noncovalent enzyme-inhibitor complex E-I. This is usually followed by a rate determining catalytic step, leading to the formation of a highly reactive species [E—I ]. This species can either undergo reaction with an active site amino acid residue of the enzyme to form the covalent enzyme-inhibitor adduct E—I", or be released into the medium to form product P and free active enzyme E. 

Several other changes that are supposed to slow down the reaction can cause runaway. In the case of ethylene oxidation, chlorinated hydrocarbons are used as inhibitors. These slow down both the total and the epoxidation, although the latter somewhat less. When the reaction is running too high and the inhibitor feed is suddenly increased in an attempt to control it, a runaway may occur. The reason is similar to that for the feed temperature cut situation. Here the inhibitor that is in the ppm region reacts with the front of the catalytic bed and slowly moves down stream. The unconverted reactants reach the hotter zone before the increased inhibitor concentration does. 

The basic kinetic properties of this allosteric enzyme are clearly explained by combining Monod s theory and these structural results. The tetrameric enzyme exists in equilibrium between a catalytically active R state and an inactive T state. There is a difference in the tertiary structure of the subunits in these two states, which is closely linked to a difference in the quaternary structure of the molecule. The substrate F6P binds preferentially to the R state, thereby shifting the equilibrium to that state. Since the mechanism is concerted, binding of one F6P to the first subunit provides an additional three subunits in the R state, hence the cooperativity of F6P binding and catalysis. ATP binds to both states, so there is no shift in the equilibrium and hence there is no cooperativity of ATP binding. The inhibitor PEP preferentially binds to the effector binding site of molecules in the T state and as a result the equilibrium is shifted to the inactive state. By contrast the activator ADP preferentially binds to the effector site of molecules in the R state and as a result shifts the equilibrium to the R state with its four available, catalytically competent, active sites per molecule. 

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