Chapter 6

Which of the following has not been shown to play a role in determining the specificity of protein kinases? A) Disulfide bonds near the phosphorylation site B) Primary sequence at phosphorylation site C) Protein quaternary structure D) Protein tertiary structure E) Residues near the phosphorylation site

Which of the following has not been shown to play a role in determining the specificity of protein kinases? A) Disulfide bonds near the phosphorylation site

Which of the following statements about allosteric control of enzymatic activity is false? A) Allosteric effectors give rise to sigmoidal V0 vs. [S] kinetic plots. B) Allosteric proteins are generally composed of several subunits. C) An effector may either inhibit or activate an enzyme. D) Binding of the effector changes the conformation of the enzyme molecule. E) Heterotropic allosteric effectors compete with substrate for binding sites.

Which of the following statements about allosteric control of enzymatic activity is false? E) Heterotropic allosteric effectors compete with substrate for binding sites.

Which of the following statements is false? A) A reaction may not occur at a detectable rate even though it has a favorable equilibrium. B) After a reaction, the enzyme involved becomes available to catalyze the reaction again. C) For S-->P, a catalyst shifts the reaction equilibrium to the right. D) Lowering the temperature of a reaction will lower the reaction rate. E) Substrate binds to an enzyme's active site.

Which of the following statements is false? C) For S-->P, a catalyst shifts the reaction equilibrium to the right.

Which of these statements about enzyme-catalyzed reactions is false? A) At saturating levels of substrate, the rate of an enzyme-catalyzed reaction is proportional to the enzyme concentration. B) If enough substrate is added, the normal Vmax of a reaction can be attained even in the presence of a competitive inhibitor. C) The rate of a reaction decreases steadily with time as substrate is depleted. D) The activation energy for the catalyzed reaction is the same as for the uncatalyzed reaction, but the equilibrium constant is more favorable in the enzyme-catalyzed reaction. E) The Michaelis-Menten constant Km equals the [S] at which V = 1/2 Vmax.

Which of these statements about enzyme-catalyzed reactions is false? D) The activation energy for the catalyzed reaction is the same as for the uncatalyzed reaction, but the equilibrium constant is more favorable in the enzyme-catalyzed reaction. *ENZYMES DO NOT CHANGE EQUILIBRIUM, ONLY REACTION RATES!*

Which one of the following statements is true of enzyme catalysts? A) Their catalytic activity is independent of pH. B) They are generally equally active on D and L isomers of a given substrate. C) They can increase the equilibrium constant for a given reaction by a thousand fold or more. D) They can increase the reaction rate for a given reaction by a thousand fold or more. E) To be effective, they must be present at the same concentration as their substrate.

Which one of the following statements is true of enzyme catalysts? D) They can increase the reaction rate for a given reaction by a thousand fold or more.

Which one of the following statements is true of enzyme catalysts? A) They bind to substrates, but are never covalently attached to substrate or product. B) They increase the equilibrium constant for a reaction, thus favoring product formation. C) They increase the stability of the product of a desired reaction by allowing ionizations, resonance, and isomerizations not normally available to substrates. D) They lower the activation energy for the conversion of substrate to product. E) To be effective they must be present at the same concentration as their substrates.

Which one of the following statements is true of enzyme catalysts? Enzymes differ from other catalysts in that only enzymes: D) They lower the activation energy for the conversion of substrate to product.

Two different enzymes are able to catalyze the same reaction, A-->B. They both have the same Vmax, but differ their Km the substrate A. For enzyme 1, the Km is 1.0 mM; for enzyme 2, the Km is 10 mM. When enzyme 1 was incubated with 0.1 mM A, it was observed that B was produced at a rate of 0.0020 mmoles/minute. a) What is the value of the Vmax of the enzymes? b) What will be the rate of production of B when enzyme 2 is incubated with 0.1 mM A? c) What will be the rate of production of B when enzyme 1 is incubated with 1 M (i.e., 1000 mM) A?

a) What is the value of the Vmax of the enzymes? a) 0.022 mmol/min b) What will be the rate of production of B when enzyme 2 is incubated with 0.1 mM A? b) 0.0022 mmol/min c) What will be the rate of production of B when enzyme 1 is incubated with 1 M (i.e., 1000 mM) A? c) 0.022 mmol/min

Define the terms "cofactor" and "coenzyme."

cofactor-any chemical component required for enzyme activity; includes both organic molecules (coenzymes) and inorganic ions coenzyme-an organic cofactor required for enzyme activity

Why does pH affect the activity of an enzyme?

pH affects the activity of an enzyme because the state of ionization of several amino acid side chains is affected by pH, and the activity of many enzymes requires that certain of the amino acid residue side chains be in a specific ionization state. (See Fig 6-20, p. 207.)

Which one of the following is not among the six internationally accepted classes of enzymes? A) Hydrolases B) Ligases C) Oxidoreductases D) Polymerases E) Transferases

(D) Polymerases are not among one of the 6 internationally accepted classes of enzymes.

Explain how a biochemist might discover that a certain enzyme is allosterically regulated.

A biochemist might discover that a certain enzyme is allosterically regulated because the enzyme would show kinetics that do not fit the Michaelis-Menten equation; the plot of V vs. [S] would be sigmoidal, not hyperbolic. The enzyme kinetics would be affected by molecules other than the substrate(s).

A good transition-state analog: A) binds covalently to the enzyme. B) binds to the enzyme more tightly than the substrate. C) binds very weakly to the enzyme. D) is too unstable to isolate. E) must be almost identical to the substrate.

A good transition-state analog: B) binds to the enzyme more tightly than the substrate.

A small molecule that decreases the activity of an enzyme by binding to a site other than the catalytic site is termed a(n): A) allosteric inhibitor. B) alternative inhibitor. C) competitive inhibitor. D) stereospecific agent. E) transition-state analog.

A small molecule that decreases the activity of an enzyme by binding to a site other than the catalytic site is termed a(n): A) allosteric inhibitor. (Heterotropic allosteric inhibitors have different specific binding sites in enzymes. Homotropic allosteric inhibitors share a regulatory and active site.)

A transition-state analog: A) is less stable when binding to an enzyme than the normal substrate. B) resembles the active site of general acid-base enzymes. C) resembles the transition-state structure of the normal enzyme-substrate complex. D) stabilizes the transition state for the normal enzyme-substrate complex. E) typically reacts more rapidly with an enzyme than the normal substrate.

A transition-state analog: C) resembles the transition-state structure of the normal enzyme-substrate complex.

What is a zymogen (proenzyme)? Explain briefly with an example.

A zymogen is an inactive form of an enzyme that is activated by one or more proteolytic cleavages in its sequence. Chymotrypsinogen, trypsinogen, and proelastase are all zymogens, becoming chymotrypsin, trypsin, and elastase, respectively, after proper cleavage.

Compare the two reaction coordinate diagrams below and select the answer that correctly describes their relationship. In each case, the single intermediate is the ES complex. A) (a) describes a strict "lock and key" model, whereas (b) describes a transition-state complementarity model. B) The activation energy for the catalyzed reaction is 5 in (a) and is 7 in (b). C) The activation energy for the uncatalyzed reaction is given by 5 + 6 in (a) and by 7 + 4 in (b). D) The contribution of binding energy is given by 5 in (a) and by 7 in (b). E) The ES complex is given by 2 in (a) and 3 in (b).

A) (a) describes a strict "lock and key" model, whereas (b) describes a transition-state complementarity model.

In the following diagram of the first step in the reaction catalyzed by the protease chymotrypsin, the process of general base catalysis is illustrated by the number ________, and the process of covalent catalysis is illustrated by the number _________. A) 1; 2 B) 1; 3 C) 2; 3 D) 2; 3 E) 3; 2

A) 1; 2

Phenyl-methane-sulfonyl-fluoride (PMSF) inactivates serine proteases by binding covalently to the catalytic serine residue at the active site; this enzyme-inhibitor bond is not cleaved by the enzyme. This is an example of what kind of inhibition? A) Irreversible B) Competitive C) Non-competitive D) Mixed E) pH inhibition

A) Irreversible

Michaelis-Menten kinetics is sometimes referred to as "saturation" kinetics. Why?

According to the Michaelis-Menten model of enzyme-substrate interaction, when [S] becomes very high, an enzyme molecule's active site will become occupied with a new substrate molecule as soon as it releases a product. Therefore, at very high [S], V0 does not increase with additional substrate, and the enzyme is said to be "saturated" with substrate.

Allosteric enzymes: A) are regulated primarily by covalent modification. B) usually catalyze several different reactions within a metabolic pathway. C) usually have more than one polypeptide chain. D) usually have only one active site. E) usually show strict Michaelis-Menten kinetics.

Allosteric enzymes: C) usually have more than one polypeptide chain.

The allosteric enzyme ATCase is regulated by CTP, which binds to the T-state of ATCase. CTP is a: A) positive regulator. B) negative regulator. C) co-factor. D) competitive inhibitor. E) coenzyme.

B) negative regulator.

Both water and glucose share an —OH that can serve as a substrate for a reaction with the terminal phosphate of ATP catalyzed by hexokinase. Glucose, however, is about a million times more reactive as a substrate than water. The best explanation is that: A) glucose has more —OH groups per molecule than does water. B) the larger glucose binds better to the enzyme; it induces a conformational change in hexokinase that brings active-site amino acids into position for catalysis. C) the —OH group of water is attached to an inhibitory H atom, while the glucose —OH group is attached to C. D) water and the second substrate, ATP, compete for the active site resulting in a competitive inhibition of the enzyme. E) water normally will not reach the active site because it is hydrophobic.

Both water and glucose share an —OH that can serve as a substrate for a reaction with the terminal phosphate of ATP catalyzed by hexokinase. Glucose, however, is about a million times more reactive as a substrate than water. The best explanation is that: B) the larger glucose binds better to the enzyme; it induces a conformational change in hexokinase that brings active-site amino acids into position for catalysis.

The following data were obtained in a study of an enzyme known to follow Michaelis-Menten kinetics: V0 Substrate added (micromol/min) (mmol/L) ————————————— 217 0.8 325 2 433 4 488 6 647 1,000 ————————————— The Km for this enzyme is approximately: A) 1 mM. B) 1000 mM. C) 2 mM. D) 4 mM. E) 6 mM.

C) 2 mM.

Michaelis and Menten assumed that the overall reaction for an enzyme-catalyzed reaction could be written as E + S (k1,k-1)> ES(k2) > E + P Using this reaction, the rate of breakdown of the enzyme-substrate complex can be described by the expression: A) k1 ([Et] - [ES]). B) k1 ([Et] - [ES])[S]. C) k2 [ES]. D) k-1 [ES] + k2 [ES]. E) k-1 [ES].

D) k-1 [ES] + k2 [ES].

For the simplified representation of an enzyme-catalyzed reaction shown below, the statement "ES is in steady-state" means that: E + S (k1,k-1) > ES (k2, k-2) > E + P A) k2 is very slow. B) k1= k2. C) k1= k-1. D) k1[E][S] = k-1[ES] + k2[ES]. E) k1[E][S] = k-1[ES].

D) k1[E][S] = k-1[ES] + k2[ES].

Blood coagulation involves: A) a kinase cascade. B) zymogen activation. C) serine proteases. D) A and B. E) B and C.

E) B and C.

Enzyme X exhibits maximum activity at pH = 6.9. X shows a fairly sharp decrease in its activity when the pH goes much lower than 6.4. One likely interpretation of this pH activity is that: A) a Glu residue on the enzyme is involved in the reaction. B) a His residue on the enzyme is involved in the reaction. C) the enzyme has a metallic cofactor. D) the enzyme is found in gastric secretions. E) the reaction relies on specific acid-base catalysis.

Enzyme X exhibits maximum activity at pH = 6.9. X shows a fairly sharp decrease in its activity when the pH goes much lower than 6.4. One likely interpretation of this pH activity is that: B) a His residue on the enzyme is involved in the reaction. (His residues are sensitive to pH due to the fact that the residue carries a pH of 6.0. Relatively small changes in pH changes the overall charge of His.)

Enzymes are potent catalysts because they: A) are consumed in the reactions they catalyze. B) are very specific and can prevent the conversion of products back to substrates. C) drive reactions to completion while other catalysts drive reactions to equilibrium. D) increase the equilibrium constants for the reactions they catalyze. E) lower the activation energy for the reactions they catalyze.

Enzymes are potent catalysts because they: E) lower the activation energy for the reactions they catalyze.

Enzymes differ from other catalysts in that only enzymes: A) are not consumed in the reaction. B) display specificity toward a single reactant. C) fail to influence the equilibrium point of the reaction. D) form an activated complex with the reactants. E) lower the activation energy of the reaction catalyzed.

Enzymes differ from other catalysts in that only enzymes: B) display specificity toward a single reactant.

Methanol (wood alcohol) is highly toxic because it is converted to formaldehyde in a reaction catalyzed by the enzyme alcohol dehydrogenase: NAD+ + methanol --> NADH + H+ + formaldehyde Part of the medical treatment for methanol poisoning is to administer ethanol (ethyl alcohol) in amounts large enough to cause intoxication under normal circumstances. Explain this in terms of what you know about examples of enzymatic reactions.

Ethanol is a structural analog of methanol, and competes with methanol for the binding site of alcohol dehydrogenase, slowing the conversion of methanol to formaldehyde, and allowing its clearance by the kidneys. The effect of ethanol is that of a competitive inhibitor

For a reaction that can take place with or without catalysis by an enzyme, what would be the effect of the enzyme on the: (a) standard free energy change of the reaction? (b) activation energy of the reaction? (c) initial velocity of the reaction? (d) equilibrium constant of the reaction?

For a reaction that can take place with or without catalysis by an enzyme, what would be the effect of the enzyme on the: (a) standard free energy change of the reaction? -no change (b) activation energy of the reaction? decrease (c) initial velocity of the reaction? increase (d) equilibrium constant of the reaction? no change

For enzymes in which the slowest (rate-limiting) step is the reaction: ES (k2) --> P Km becomes equivalent to: A) kcat. B) the [S] where V0 = Vmax. C) the dissociation constant, Kd, for the ES complex. D) the maximal velocity. E) the turnover number.

For enzymes in which the slowest (rate-limiting) step is the reaction: k2 ES --> P Km becomes equivalent to: C) the dissociation constant, Kd, for the ES complex.

The enzymatic activity of lysozyme is optimal at pH 5.2 and decreases above and below this pH value. Lysozyme contains two amino acid residues in the active site essential for catalysis: Glu35 and Asp52. The pK value for the carboxyl side chains of these two residues are 5.9 and 4.5, respectively. What is the ionization state of each residue at the pH optimum of lysozyme? How can the ionization states of these two amino acid residues explain the pH-activity profile of lysozyme?

For the enzyme to be active, it is likely that Asp52 is unprotonated and Glu35 is protonated. When the pH is below 4.5, Asp52 becomes protonated, and when it is above 5.9, Glu35 is deprotonated, either of which decreases the activity of the enzyme. (See Fig. 6-20, p. 207.)

In a plot of l/V against 1/[S] for an enzyme-catalyzed reaction, the presence of a competitive inhibitor will alter the: A) curvature of the plot. B) intercept on the l/[S] axis. C) intercept on the l/V axis. D) pK of the plot. E) Vmax.

In a plot of l/V against 1/[S] for an enzyme-catalyzed reaction, the presence of a competitive inhibitor will alter the: B) intercept on the l/[S] axis. (Because competitive inhibitors compete with [S]).

Chymotrypsin belongs to a group of proteolyticenzymes called the "serine proteases," many of which have an Asp, His, and Ser residue that are crucial to the catalytic mechanism. The serine hydroxyl functions as a nucleophile. What do the other two amino acids do to support this nucleophilic reaction?

In chymotrypsin, histidine functions as a general base, accepting a proton from the serine hydroxyl, thereby increasing serine's reactivity as a nucleophile. The negatively charged Asp stabilizes the positive charge that develops on the His.

In competitive inhibition, an inhibitor: A) binds at several different sites on an enzyme. B) binds covalently to the enzyme. C) binds only to the ES complex. D) binds reversibly at the active site. E) lowers the characteristic Vmax of the enzyme.

In competitive inhibition, an inhibitor: D) binds reversibly at the active site.

An enzyme-catalyzed reaction was carried out with the substrate concentration initially a thousand times greater than the Km for that substrate. After 9 minutes, 1% of the substrate had been converted to product, and the amount of product formed in the reaction mixture was 12 μmol. If, in a separate experiment, one-third as much enzyme and twice as much substrate had been combined, how long would it take for the same amount (12 μmol) of product to be formed? A) 1.5 min B) 13.5 min C) 27 min D) 3 min E) 6 min

It would take 27 minutes for the same amount of product to be formed. (Think about it: The first time it takes 9 minutes for only 1% to be formed. Therefore, if the amount of substrate is twice the amount as the previous but there was 1/3 less of the amount of enzyme, then it would take about 3 times as long for the reduced amount of enzyme to bind to the substrate. Even though there is more substrate, there is 3 times less enzyme and the rate of an enzyme-catalyzed reaction is proportional to the amount of [E].)

Write an equilibrium expression for the reaction S --> P and briefly explain the relationship between the value of the equilibrium constant and free energy.

Keq' = [P]/[S]. The value of Keq' reflects the difference between the free energy content of S and P. Free energy and equilibrium constant are related by the expression: delta G'° = -RT ln Keq' For each change in Keq' by one order of magnitude, delta G'° changes by 5.7 Kjoule/mole.

Penicillin and related drugs inhibit the enzyme ________; this enzyme is produced by _________. A) β-lacamase; bacteria B) transpeptidase; human cells C) transpeptidase; bacteria D) lysozyme; human cells E) aldolase; bacteria

Penicillin and related drugs inhibit the enzyme _______; this enzyme is produced by _______. C) transpeptidase; bacteria

What is the difference between general acid-base catalysis and specific acid-base catalysis? (Assume that the solvent is water.)

Specific acid-base catalysis refers to catalysis by the constituents of water, (i.e., the donation of a proton by the hydronium ion, H3O+ or the acceptance of a proton by the hydroxyl ion OH-.) General acid-base catalysis refers to the donation or acceptance of a proton by weak acids and bases other than water.

The Lineweaver-Burk plot is used to: A) determine the equilibrium constant for an enzymatic reaction. B) extrapolate for the value of reaction rate at infinite enzyme concentration. C) illustrate the effect of temperature on an enzymatic reaction. D) solve, graphically, for the rate of an enzymatic reaction at infinite substrate concentration. E) solve, graphically, for the ratio of products to reactants for any starting substrate concentration.

The Lineweaver-Burk plot is used to: D) solve, graphically, for the rate of an enzymatic reaction at infinite substrate concentration. (Remember, Vmax is how much substrate is necessary for an enzymatic reaction to occur at its maximum rate, and the Lineweaver-Burk plot measures 1/Km and 1/[S] to get enzymatic rates.)

Give the Michaelis-Menten equation and define each term in it. Does this equation apply to all enzymes? If not, to which kind does it not apply?

The Michaelis-Menten equation is: V0 = Vmax [S]/( Km + [S]) -V0 is the initial velocity at any given concentration of S. -Vmax is the velocity when all enzyme molecules are saturated with S. -[S] is the concentration of S. -Km is a constant characteristic for the enzyme. This equation does not apply to enzymes that display sigmoidal V0 vs. [S] curves, but only to those giving hyperbolic kinetic plots.

The benefit of measuring the initial rate of a reaction V0 is that at the beginning of a reaction: A) [ES] can be measured accurately. B) changes in [S] are negligible, so [S] can be treated as a constant. C) changes in Km are negligible, so Km can be treated as a constant. D) V0 = Vmax. E) varying [S] has no effect on V0.

The benefit of measuring the initial rate of a reaction V0 is that at the beginning of a reaction: (B) changes in [S] are negligible, so [S] can be treated as a constant. ([S] is normally far greater than [Et] (the sum of free and substrate-bound enzyme), so the amount of substrate bound at any given time is negligible compared with the total [S].)

Which of the following is true of the binding energy derived from enzyme-substrate interactions? A) It cannot provide enough energy to explain the large rate accelerations brought about by enzymes. B) It is sometimes used to hold two substrates in the optimal orientation for reaction. C) It is the result of covalent bonds formed between enzyme and substrate. D) Most of it is derived from covalent bonds between enzyme and substrate. E) Most of it is used up simply binding the substrate to the enzyme.

The binding energy derived from enzyme-substrate interactions is B) sometimes used to hold two substrates in the optimal orientation for reaction.

On the enzyme hexokinase, ATP reacts with glucose to produce glucose 6-phosphate and ADP five orders of magnitude faster than ATP reacts with H2O to form phosphate and ADP. The intrinsic chemical reactivity of the —OH group in water is about the same as that of the glucose molecule, and water can certainly fit into the active site. Explain this rate differential in two sentences or less.

The binding of glucose to hexokinase induces a conformation change that brings the amino acid residues that facilitate the phosphoryl transfer into position in the active site. Binding of water alone does not induce this conformational change.

The concept of "induced fit" refers to the fact that: A) enzyme specificity is induced by enzyme-substrate binding. B) enzyme-substrate binding induces an increase in the reaction entropy, thereby catalyzing the reaction. C) enzyme-substrate binding induces movement along the reaction coordinate to the transition state. D) substrate binding may induce a conformational change in the enzyme, which then brings catalytic groups into proper orientation. E) when a substrate binds to an enzyme, the enzyme induces a loss of water (desolvation) from the substrate.

The concept of "induced fit" refers to the fact that: D) substrate binding may induce a conformational change in the enzyme, which then brings catalytic groups into proper orientation.

The difference in (standard) free energy content, delta G'°, between substrate S and product P may vary considerably among different reactions. What is the significance of these differences?

The difference in free energy content between substrate (or reactant) and product for each reaction reflects the relative amounts of each compound present at equilibrium. The greater the difference in free energy, the greater the difference in amounts of each compound at equilibrium.

The double-reciprocal transformation of the Michaelis-Menten equation, also called the Lineweaver-Burk plot, is given by 1/V0 = Km /(Vmax[S]) + 1/Vmax. To determine Km from a double-reciprocal plot, you would: A) multiply the reciprocal of the x-axis intercept by -1. B) multiply the reciprocal of the y-axis intercept by -1. C) take the reciprocal of the x-axis intercept. D) take the reciprocal of the y-axis intercept. E) take the x-axis intercept where V0 = 1/2 Vmax.

The double-reciprocal transformation of the Michaelis-Menten equation, also called the Lineweaver-Burk plot, is given by 1/V0 = Km /(Vmax[S]) + 1/Vmax. To determine Km from a double-reciprocal plot, you would: A) multiply the reciprocal of the x-axis intercept by -1.

A metabolic pathway proceeds according to the scheme, R --> S --> T --> U --> V --> W. A regulatory enzyme, X, catalyzes the first reaction in the pathway. Which of the following is most likely correct for this pathway? A) Either metabolite U or V is likely to be a positive modulator, increasing the activity of X. B) The first product S, is probably the primary negative modulator of X, leading to feedback inhibition. C) The last product, W, is likely to be a negative modulator of X, leading to feedback inhibition. D) The last product, W, is likely to be a positive modulator, increasing the activity of X. E) The last reaction will be catalyzed by a second regulatory enzyme.

The most likely correct for the pathway R-->S-->T-->U-->V-->W is: C) The last product, W, is likely to be a negative modulator of X, leading to feedback inhibition.

The number of substrate molecules converted to product in a given unit of time by a single enzyme molecule at saturation is referred to as the: A) dissociation constant. B) half-saturation constant. C) maximum velocity. D) Michaelis-Menten number. E) turnover number.

The number of substrate molecules converted to product in a given unit of time by a single enzyme molecule at saturation is referred to as the: E) turnover number.

Why is the Lineweaver-Burk (double reciprocal) plot (see Box 6, p. 206) more useful than the standard V vs. [S] plot in determining kinetic constants for an enzyme? (Your answer should probably show typical plots.)

The plot of V vs. [S] is hyperbolic; maximum velocity is never achieved experimentally, because it is impossible to do experiments at infinitely high [S]. The Lineweaver-Burk transformation of the Michaelis-Menten equation produces a linear plot that can be extrapolated to infinite [S] (where 1/[S] becomes zero), allowing a determination of Vmax.

Sometimes the difference in (standard) free-energy content, delta G'°, between a substrate S and a product P is very large, yet the rate of chemical conversion, S --> P, is quite slow. Why?

The rate of conversion from substrate to product (or the reverse reaction, from product to substrate) does not depend on the free-energy difference between them. The rate of the reaction depends upon the activation energy of the reaction delta G'‡, which is the difference between the free-energy content of S (or P) and the reaction transition state.

The role of an enzyme in an enzyme-catalyzed reaction is to: A) bind a transition state intermediate, such that it cannot be converted back to substrate. B) ensure that all of the substrate is converted to product. C) ensure that the product is more stable than the substrate. D) increase the rate at which substrate is converted into product. E) make the free-energy change for the reaction more favorable.

The role of an enzyme in an enzyme-catalyzed reaction is to: D) increase the rate at which substrate is converted into product.

The role of the metal ion (Mg2+) in catalysis by enolase is to: A) act as a general acid catalyst B) act as a general base catalyst C) facilitate general acid catalysis D) facilitate general base catalysis E) stabilize protein conformation

The role of the metal ion (Mg2+) in catalysis by enolase is to: D) facilitate general base catalysis

For serine to work effectively as a nucleophile in covalent catalysis in chymotrypsin a nearby amino acid, histidine, must serve as general base catalyst. Briefly describe, in words, how these two amino acids work together.

The serine is a polar hydroxyl, with the oxygen functioning as an electronegative nucleophile. A nearby histidine residue, with pKa 6.0, however, functions as a base to abstract the proton from the serine hydroxyl group. The result is to substantially increase the electronegativity of the serine oxygen, making it a much stronger nucleophile. This, in turn, lowers the activation energy of the covalent catalysis between serine and the carbonyl carbon of the substrate peptide bond.

Which of the following statements about a plot of V0 vs. [S] for an enzyme that follows Michaelis-Menten kinetics is false? A) As [S] increases, the initial velocity of reaction V0 also increases. B) At very high [S], the velocity curve becomes a horizontal line that intersects the y-axis at Km. C) Km is the [S] at which V0 = 1/2 Vmax. D) The shape of the curve is a hyperbola. E) The y-axis is a rate term with units of μm/min.

The statement that is false concerning a plot of V0 vs. [S] for an enzyme that follows Michaelis-Menten kinetics is: (B) At very high [S], the velocity curve becomes a horizontal line that intersects the y-axis at Km. -At very high [S], the velocity curve becomes a plateau-like and is close to to Vmax.

The steady state assumption, as applied to enzyme kinetics, implies: A) Km = Ks. B) the enzyme is regulated. C) the ES complex is formed and broken down at equivalent rates. D) the Km is equivalent to the cellular substrate concentration. E) the maximum velocity occurs when the enzyme is saturated.

The steady state assumption, as applied to enzyme kinetics, implies: C) the ES complex is formed and broken down at equivalent rates.

Penicillin and related antibiotics contain a 4-membered Beta-lactam ring. Explain why this feature is important to the mechanism of action of these drugs.

The strained 4-membered ring is easily opened (deltaG << 0); this energy drives the reaction that covalently inactivates the transpeptidase.

Why is a transition-state analog not necessarily the same as a competitive inhibitor?

The structure of a competitive inhibitor may be similar to the structure of the free substrate. Similar structure will mean that the competitive inhibitor can associate with the enzyme at the active site, effectively blocking the normal substrate from binding. A transition-state analog, however, is similar in structure to the transition-state of the reaction catalyzed by the enzyme. Often a transition-state analog will bind tightly to an enzyme, and is not easily competed away by substrate.

How does the total enzyme concentration affect turnover number and Vmax?

The turnover number, kcat, is the number of substrate molecules converted to product in a given time by a single enzyme molecule, so turnover number is not affected by the total enzyme concentration, [Et]. For any given reaction, however, Vmax can change because Vmax is the product of turnover number x the total enzyme concentration, or Vmax = kcat [Et].

To calculate the turnover number of an enzyme, you need to know: A) the enzyme concentration. B) the initial velocity of the catalyzed reaction at [S] >> Km. C) the initial velocity of the catalyzed reaction at low [S]. D) the Km for the substrate. E) both A and B.

To calculate the turnover number of an enzyme, you need to know: E) both A and B, which are: A) the enzyme concentration. B) the initial velocity of the catalyzed reaction at [S] >> Km.

How is trypsinogen converted to trypsin? A) A protein kinase-catalyzed phosphorylation converts trypsinogen to trypsin. B) An increase in Ca2+ concentration promotes the conversion. C) Proteolysis of trypsinogen forms trypsin. D) Trypsinogen dimers bind an allosteric modulator, cAMP, causing dissociation into active trypsin monomers. E) Two inactive trypsinogen dimers pair to form an active trypsin tetramer.

Trypsinogen is converted to trypsin by: C) Proteolysis of trypsinogen forms trypsin.

One of the enzymes involved in glycolysis, aldolase, requires Zn2+ for catalysis. Under conditions of zinc deficiency, when the enzyme may lack zinc, it would be referred to as the: A) apoenzyme. B) coenzyme. C) holoenzyme. D) prosthetic group. E) substrate.

Under conditions of zinc deficiency, when the enzyme may lack zinc, it would be referred to as the: A) apoenzyme. (holoenzyme=enzyme + coenzyme)

An enzyme can catalyze a reaction with either of two substrates, S1 or S2. The Km for S1 was found to be 2.0 mM, and the Km, for S2 was found to be 20 mM. A student determined that the Vmax was the same for the two substrates. Unfortunately, he lost the page of his notebook and needed to know the value of Vmax. He carried out two reactions: one with 0.1 mM S1, the other with 0.1 mM S2. Unfortunately, he forgot to label which reaction tube contained which substrate. Determine the value of Vmax from the results he obtained: Tube number Rate of formation of product 1 0.5 2 4.8

Vmax = 101

Vmax for an enzyme-catalyzed reaction: A) generally increases when pH increases. B) increases in the presence of a competitive inhibitor. C) is limited only by the amount of substrate supplied. D) is twice the rate observed when the concentration of substrate is equal to the Km. E) is unchanged in the presence of a uncompetitive inhibitor.

Vmax for an enzyme-catalyzed reaction: D) is twice the rate observed when the concentration of substrate is equal to the Km. ([S] and therefore Km are increased, but Vmax stays the same.)