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Competitive Inhibition That Barrier, As We Shall See

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Competitive Inhibition That Barrier, As We Shall See Chapter 4 Catalysis In living systems, speed is everything. Providing the reaction speeds necessary to support life are the catalysts, mostly in the form of enzymes. Catalysis Introduction Introduction Activation Energy General Mechanisms of Action If there is a magical component to life, an argument can surely be made for it Substrate Binding being catalysis. Thanks to catalysis, reactions that could take hundreds of years Enzyme Flexibility to complete in the “real world,” occur in seconds in the presence of a catalyst. Active Site Chymotrypsin Chemical catalysts, like platinum, speed reactions, but enzymes (which are simply Enzyme Parameters super-catalysts with a twist) put chemical catalysts to shame. To understand VMAX & KCAT enzymatic catalysis, we must first understand energy. In Chapter 2, we noted the KM tendency for processes to move in the direction of lower energy. Chemical Perfect Enzymes Lineweaver-Burk Plots reactions follow this universal trend, but they often have a barrier in place that Enzyme Inhibition must be overcome. The secret to catalytic action is reducing the magnitude of Competitive Inhibition that barrier, as we shall see. No Effect On VMAX Increased KM Non-Competitive Inhibition Uncompetitive Inhibition Suicide Inhibition Control of Enzymes Allosterism Covalent Control of Enzymes Other Controls of Enzymes Ribozymes Enzyme Rate Enhancements 81 Activation Energy Another feature to note about catalyzed reactions is Click HERE, HERE, and HERE for The figure below schematically depicts the energy changes that the reduced energy barrier Kevin’s Catalytic Mechanism occur during the progression of a simple reaction. In the figure, lectures on YouTube (also called the activation the energy differences during the reaction are compared for a energy or free energy of catalyzed (plot on the right) and an uncatalyzed reaction (plot on activation) to reach the transition state of the catalyzed reaction. the left). Notice that the reactants start at the same energy level This is the second important point for both conditions and about catalyzed reactions – catalysts that the products end work by lowering activation energies at the same energy for of reactions and thus molecules more both as well. Thus, the easily reach the energy necessary to difference in energy get to the point where the reaction between the energy of occurs. Note that these reactions are the ending compounds reversible. The extent to which they and the starting will proceed is a function of the size compounds is the same of the energy difference between the in both cases. This is product and reactant states. The the first important rule lower the energy of the products to understand any kind compared to the reactants, the larger of catalysis – catalysts the percentage of molecules that will do not change the be present as products at equilibrium. overall energy of a At equilibrium, of course, no change reaction. Given enough in concentration of reactants and time, a non-catalyzed products occurs because at this reaction will get to the point, the forward and reverse same equilibrium as a Energetic Considerations of Catalysis reaction rates are the same. catalyzed one. 82 General Mechanisms of Action Substrate Binding As noted above, enzymes are orders of magnitude more effective Another important difference between the mechanism of action of (faster) than chemical catalysts. The secret of their success lies an enzyme and a chemical catalyst is that an enzyme has binding in a fundamental difference in their mechanisms of action. Every sites that not only ‘grab’ the substrate (molecule involved in the chemistry student has had hammered into their heads the fact reaction being catalyzed), but also place it in a position to be that a catalyst speeds a reaction without being consumed by it. electronically induced to react, either within itself or with another In other words, the catalyst ends up after a reaction just the way it substrate. The enzyme itself may play a role in the electronic started so it can catalyze induction or the induction may occur as a result of substrates other reactions, as well. being placed in very close proximity to each other. Chemical Click HERE, HERE, HERE, and Enzymes share this catalysts have no such ability to bind substrates and are HERE for Kevin’s enzyme property, but in the middle, dependent upon them colliding in the right orientation at or near catalysis lectures on YouTube during the catalytic action, their surfaces. an enzyme is transiently changed. Such changes may be subtle electronic ones or more significant covalent modifications. It is also important to recognize that enzymes are not fixed, rigid structures, but rather are flexible. Flexibility allows movement and movement facilitates alteration of electronic environments necessary for catalysis. Enzymes are, thus, much more efficient than rigid chemical catalysts as a result of their abilities to facilitate the changes necessary to optimize the catalytic process. Mechanism of Induced Fit 83 Enzyme Flexibility The Way They Work To the tune of “The Way We Were” As mentioned earlier, a difference between an enzyme and a chemical catalyst is that an enzyme is flexible. Its slight Enzymes Mighty powerhouse peptides changes in shape (often arising from the binding of the Cause reactions to go faster In the cell’s insides substrate itself) help to position substrates for reaction Tiny substrates after they bind. These changes in shape are explained, in Bring about an induced fit part, by Koshland’s Induced Fit Model of Catalysis, Enzyme structure is affect-ed By what binds to it which illustrates that not only do enzymes change Can it be that it’s just simple zen? substrates, but that substrates also transiently change How the enzymes activate enzymes. At the end of the catalysis, the enzyme is If they bind effector, will they go To an R-State, T-State? returned to its original state. Enzyme flexibility also is Folding important for control of enzyme activity. Two distinct Gives the mechanistic might structures are typically described– the T (tight) state, which To three-D arrangement Of the active site is a lower activity state and the R (relaxed) state, which has Enzymes greater activity. Have a bias they can’t hide Hydrophobic side chains are Mostly found inside Active Site So it’s the structure Reactions in enzymes are catalyzed at a specific location For celebrating Whenever there’s debating known as the ‘active site’. Substrate binding sites are The way they work located in close physical proximity to the active site and oriented to provide access for the relevant portion of the T molecule to the electronic environment of the enzyme where catalysis is initiated. Recorded by Liz Bacon and David Simmons Lyrics by Kevin Ahern 84 Chymotrypsin two phases – a fast phase that occurs first and a slower phase Consider the mechanism that follows. The enzyme has a of catalysis of the enzyme substrate binding site that known as chymotrypsin. includes a region of the enzyme Found in our digestive known as the S1 pocket. Let us system, chymotrypsin’s step through the mechanism by catalytic action is cleaving which chymotrypsin cuts peptide bonds in proteins adjacent to phenylalanine. and it uses the side chain of a serine in its The process starts with the mechanism of catalysis. binding of the substrate in the Many other protein- S1 pocket. The S1 pocket in cutting enzymes employ a chymotrypsin has a hydrophobic very similar mechanism hole in which the substrate is and they are known bound. Preferred substrates will collectively as serine include amino acid side chains proteases. As a that are hydrophobic, like protease, it acts fairly phenylalanine. If an ionized side specifically, cutting not all chain, like that of glutamic acid peptide bonds, but only binds in the S1 pocket, it will those that are adjacent to quickly exit, much like water specific amino acids in the would avoid an oily interior. protein. One of the amino When the proper substrate acids it cuts adjacent to is binds, it stays and its presence phenylalanine. The induces an ever so slight shift in enzyme’s action occurs in Serine Protease Mechanism the shape of the enzyme. This 85 subtle shape change on the binding of the proper substrate starts The Serine Protease Song the steps of the catalysis and is the reason that the enzyme To the tune of “Blackbird” shows specificity for cutting at specific enzyme positions in the Substrate floating in the cell’s insides Enzyme snags it with its binding site target protein. Only amino acids with the side chains that interact It supplies well with the S1 pocket start the catalytic wheels turning. Shuffling of electrons in the act to catalyze The slight changes in shape of the enzyme upon binding of the Proteases of the serine kind Break up peptide bonds in rapid time proper substrate cause changes in the positioning of three amino Fast and slow acids (aspartic acid, histidine, and serine) in the active site known Steps in breaking bonds are mechanisms you should know as the catalytic triad, during the second step of the catalytic Asp – his - ser action. The shift of the negatively charged aspartic acid towards Bonds beware the electron rich histidine ring favors the abstraction of a proton Inside the S1 pocket substrate sits by the histidine from the hydroxyl group on the side chain of Alkoxides serine, resulting in production of a very reactive alkoxide ion in Break peptides Nucleophiles give bonds the fits the active site. Since the active site at this point also contains the polypeptide chain positioned with the phenylalanine side chain Peptide one exits easily But water has to let the other flee embedded in the S1 pocket, the alkoxide ion performs a Bound not free nucleophilic attack on the peptide bond on the carboxyl side of ‘Cuz the enzyme’s linked to it in mechanism three phenylalanine sitting in the active site.
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