Properties of Enzymes Enzymes are catalysts What properties would ideal catalysts have? Enzymes are catalysts What properties would ideal catalysts have? 1. High degree of specificity for their substrates. 2. Accelerate chemical reactions tremendously. 3. Function in mild conditions. 4. Be recycled to participate again. A Few Definitions Cofactor – additional chemical component needed for catalysis. - often an inorganic metal ion (mineral). Coenzyme – complex organic molecule needed for catalysis. - often a vitamin. Prosthetic group – non amino acid portion of the enzyme needed for catalysis. Often a coenzyme or metal ion. Holoenzyme – complete catalytically active enzyme, with all necessary prosthetic groups. Apoenzyme – The protein part of the holoenzyme. Prosthetic groups are absent. Six Classes of Enzymes 1. Oxidoreductases 2. Transferases 3. Hydrolases 4. Lyases 5. Isomerases 6. Ligases One bond to oxygen Two bonds to oxygen Stored electrons Amino group transferred Water did chemistry to break a bond CO2 was removed 5. 5. Amino group switched places 6. 6. Two substrates were ligated together Enzyme Kinetics E = enzyme S = substrate P = product ES = enzyme-substrate complex k = rate constant The rate of reaction is dependent on enzyme concentration [Enzyme] <<< [substrate] Figure 5.2 Velocity, or how fast the reaction is going Concentration of enzyme When measuring rates of enzyme-catalyzed reactions, initial velocity (vo) is measured. Figure 5.3 Enzyme kinetics terminology [S] – substrate concentration Vo – initial velocity of a reaction. A significant amount of substrate has not yet been converted to product. Vmax – maximal velocity of a reaction. Addition of more substrate will not increase the rate of the reaction. Km – The concentration of substrate at which the rate of the reaction is half-maximal Michaelis-Menten equation Page 141 The rate of reaction is dependent on substrate concentration Figure 5.4 Km values are often just above the substrate concentration in a cell. Rates of reaction are sensitive to small changes in cellular substrate concentrations. kcat is a measure of the number of substrate molecules converted to product per second per enzyme molecule Experimental determination of kinetic constants Figure 5.6 Reversible Enzyme Inhibition An inhibitor is a compound that binds to an enzyme and interferes with its activity. Many drugs are enzyme inhibitors. An inhibitor is characterized by an inhibition constant (Ki). No Inhibitor Figures 5.8 and 5.9 No Inhibitor Many competitive inhibitors are substrate analogs Benzamidine is an inhibitor of the enzyme trypsin Many competitive inhibitors are substrate analogs. Compound (b) designed as an inhibitor of the enzyme purine nucleoside phosphorylase, that utilizes guanosine (a) as a substrate. (b) is a possible drug for the treatment of arthritis. Figure 5.13 Irreversible Enzyme Inhibition Some inhibitors are compounds that form a stable covalent bond with the target enzyme. DFP inactivates serine proteases by covalently modifying an active site serine residue. Figure 5.15 F- Regulation of enzyme activity by metabolite concentration Activator Regulator ADP binds at an allosteric site, Separate from the active site The activity of the some enzymes is regulated by reversible phosphorylation. Example: pyruvate dehydrogenase Figure 5.22