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2. Taxol is an anti-cancer drug that blocks depolymerization of microtubules, this could result
in:
A. prevention of tyrosine kinase receptor dimerization
B. inhibition of chromosomal partitioning during mitosis
C. inhibition of filopodial extensions at the leading edge of metastatic cells
D. impaired lamin structure underlying the nuclear membrane
E. cell lysis
3. An uncatalyzed reaction has a rate of 4.2 x 10–7 s–1. When an enzyme is added the rate is 3.2
x 104 s–1. Calculate the rate enhancement caused by the enzyme.
A. 7.6 x 1010
B. 3.2 x 104
C. 1.3 x 10–2
D. 7.4 x 10–3
E. cannot be determined
1
BIOC 2300 Week 4 Additional Practice Questions Feb 8, 2017
5. An enzyme that forms a covalent bond with its substrate during the course of a reaction is
considered to undergo _____.
A. acid-base catalysis
B. electrophilic catalysis
C. covalent catalysis
D. metal ion catalysis
E. none of the above
6. The ability for an enzyme to change its shape upon substrate binding represents the concept of
_____.
A. lock and key
B. induced fit
C. proximity and orientation effects
D. covalent catalysis
E. none of the above
7. When a substrate and enzyme interact, the first chemical species formed is the _____.
A. enzyme-substrate complex
B. enzyme-transition state complex
C. enzyme-product complex
D. enzyme plus product
E. none of the above
8. When the Michaelis constant (KM) is considered to be very similar to the dissociation constant
for the ES complex, then:
A. ES → E + P is fast compared to ES → E + S.
B. the turnover number is very large.
C. kcat/KM is near the diffusion-controlled limit.
D. k2 << k-1
E. the enzyme has been inhibited.
2
BIOC 2300 Week 4 Additional Practice Questions Feb 8, 2017
12. The protein kinase Raf is a key enzyme in growth factor signaling pathways leading to cell
proliferation, and thus a potential anti-cancer drug target. In screening natural products for
inhibitors of Raf, your drug company identifies a compound (DMB12345) that has the
following effects on the phosphorylation of a peptide substrate by ATP, when enzyme kinetic
data are displayed using a double-reciprocal plot:
1 / [Vo (µmol/min)]
+ DMB12345
4
2 - DMB12345
1 2 3
3
1 / [ATP (µM)]
BIOC 2300 Week 4 Additional Practice Questions Feb 8, 2017
13. Why will the KM of an enzyme for a particular substrate always be greater than the
dissociation constant (KD) of the enzyme for that substrate?
14. Radioactively labelled [14C]-proline is incorporated into collagen in cultured fibroblast cells.
The radioactivity is detected in the collagen protein. However, collagen synthesized in the
presence of [14C]-hydroxyproline is not labeled. Explain why.
15. Most human enzymes have narrow temperature optima around 37˚C. Explain why.
16. Bacterial maltase catalyzes the hydrolysis of the disaccharide maltose to two glucose
molecules. The enzyme has a KM for maltose of 0.135 µM and a VMAX of 65 µmol/min.
What is the reaction velocity when the concentration of maltose is 1 µM?
17. When [S] = 5 x KM, how close is v0 to VMAX? When [S] = 20 x KM, how close is v0 to
VMAX? What do these results tell you about the accuracy of estimating VMAX from a plot of
v0 versus [S]?
4
BIOC 2300 Week 4 Additional Practice Questions Feb 8, 2017
Answers
1. A
2. B
4. E
5. C
6. B
7. A
9. D (answer obtained by plugging values into the MM equation and solving for Km)
10. A
11. B
12. B
13. For the simple enzyme reaction E + S <--> ES ---> E + P, the KM is defined as (k-1 + k2)/k1,
whereas the dissociation constant (KD) for the ES complex back to E + S is k-1/ k1. Thus, KM
must always greater than KD, and only when k2 is much less than k-1 does KM approach KD.
(Note this is also the answer to question 8)
14. The individual collagen chains are synthesized on the ribosome from the constituent amino
acids. Proline is one of the 20 amino acids for which a codon exists in the mRNA. Some of
the proline residues are modified post-translationally (after the protein is synthesized) to
hydroxyproline. Thus, hydroxyproline itself is not incorporated directly into the protein
during synthesis (there is no codon for hydroxyproline) and so no radioactivity from [14C]-
hydroxyproline would appear in the collagen product.
15. At first, the temperature increases the reaction rate because heat generally increases the
proportion of reacting groups that can achieve the transition state in a given time. However,
when the temperature rises above a certain point, the heat causes the enzyme, which is a
protein, to denature. This accounts for the steep drop in enzymatic activity at higher
temperatures.
5
BIOC 2300 Week 4 Additional Practice Questions Feb 8, 2017
Similarly, when [S] = 20 KM, the velocity is 95% of the maximum velocity. Therefore, a four-
fold increase in substrate concentration causes a smaller proportional increase in velocity (from
83% to 95%). Estimating Vmax from a plot of v0 versus [S] is difficult because the substrate
concentration must be quite high in order to achieve a maximal velocity of close to 100%. It is
possible that these points on the hyperbolic curve cannot be experimentally measured since at
high concentration the substrate may not be soluble in the reaction medium. It is better to obtain
Vmax by fitting experimental data to the equation of a hyperbola or, alternatively, from a
Lineweaver–Burk plot.