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Enzyme Mechanism and Chemical Equation Questionnaire
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I’m stuck on a Biology question and need an explanation.
1) Consider the enzyme mechanism represented by the following chemical equation and then answer the multiple choice questions that follow:
- Which of the following is kcat (aka, the “turnover number”)?
A) k2
B) k1
C) k-1
D) k-1/k1
E) k1/k-1 - Which is NOT an accurate description of the steady state approximation?
A) d[ES]/dt = 0
B) k1[E][S] = k-1[ES] + k2[ES]
C) d[P]/dt
D) the rate of ES complex formation and break down are equivalent - Which statement regarding KM is NOT true?
A) It is defined as (k-1 + k2)/k1
B) It is equal to [E][S]/ES] (KD), if k-1 >> k2
C) It is the substrate concentration at 0.5 Vmax
D) It has units of concentration/unit time (e.g., Molar/sec) - Which of the following is NOT equivalent to Vmax during steady state conditions?
A) V0 as [S] approaches infinity
B) KM/2
C) k2[E]T
D) d[P]/dt, [S] >> KM
2) The following kinetics data were obtained for an enzyme:
[S] (μM) |
V0 (no inhibitor) (μmol/min) |
V0 (25 μM inhibitor X) (μmol/min) |
V0 (25 μM inhibitor Y) (μmol/min) |
3 |
10.4 |
4.1 |
2.1 |
5 |
14.5 |
6.4 |
2.9 |
10 |
22.5 |
11.3 |
4.5 |
30 |
33.8 |
22.6 |
6.8 |
90 |
40.5 |
33.8 |
8.1 |
- Plot V0 against [S] for these data (use graph paper or an Excel spreadsheet).
- Plot 1/V0 against 1/[S] for these data.
- What are the KM values of the enzyme without and with inhibitors?
- What are the Vmax values for the enzyme without and with inhibitors?
- What kind of inhibitor is X?
- What kind of inhibitor is Y?
- The enzyme prepartation used in the above experiment was only 15% pure. What is the Vmax of the pure enzyme?
- What is the KM of the pure enzyme?