Chapter 12: Q10CP (page 382)
What distinguishes an inhibitor from an inactivator?
Short Answer
Inhibitors reduce the activity of an enzyme while inactivators permanently block the activity of an enzyme.
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Chapter 12: Q10CP (page 382)
What distinguishes an inhibitor from an inactivator?
Inhibitors reduce the activity of an enzyme while inactivators permanently block the activity of an enzyme.
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Estimate KI for a competitive inhibitor when [I] = 5mM gives an apparent value of KM that is three times the KM for the uninhibited reaction.
Sphingosine-1-phosphate (SIP) is important for cell survival. The synthesis of SIP from sphingosine and ATP is catalyzed by the enzyme sphingosine kinase. An understanding of the kinetics of the sphingosine kinase reaction may be important in the development of drugs to treat cancer. The velocity of the sphingosine kinase reaction was measured in the presence and absence of threo-sphingosine, a stereoisomer of sphingosine that inhibits the enzyme. The results are shown below.
[Sphingosine] (饾泹惭) | v鈧 (mg min鈦宦) (no inhibitor) | v鈧 (mg min鈦宦) (with threo-sphingosine) |
2.5 | 32.3 | 8.5 |
3.5 | 40 | 11.5 |
5 | 50.8 | 14.6 |
10 | 72 | 25.4 |
20 | 87.7 | 43.9 |
50 | 115.4 | 70.8 |
Construct a Lineweaver-Burk plot to answer the following questions:
(a) What are the apparent KM and Vmax values in the presence and absence of the inhibitor?
(b) What kind of an inhibitor is threo-sphingosine? Explain.
For an enzyme-catalyzed reaction, the presence of 5 nM of a reversible inhibitor yields a Vmax value that is 80% of the value in the absenceof the inhibitor. The KM value is unchanged. (a) What type of inhibition is likely occurring? (b) What proportion of the enzyme molecules have bound inhibitor? (c) Calculate the inhibition constant.
You are attempting to determine KM by measuring the reaction velocity at different substrate concentrations, but you do not realize thatthe substrate tends to precipitate under the experimental conditions youhave chosen. How would this affect your measurement of KM?
List some advantages of phosphorylation/ dephosphorylation cascade systems over simple allosteric regulation.
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