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d. What type of inhibition is exhibited against NAD* as a cofactor? Describe what is going on in this type of inhibition. e. Referring to the mechanism below, what step is pyrazole likely interfering with given your answers in c and d? 1. binding of NAD+ to the enzyme, LADH 2. binding of C₂H5OH to LADH 3. Oxidation of C₂H5OH (This is the reaction step) and product dissociation 4. Dissociation of NADH from LADH NAD + LADH NAD LADH + C₂H5OH NAD LADH C₂H5OH NADH LADH NAD LADH k2 NAD LADH C₂H5OH ОН NADH LADH + NADH+ LADH CH3CHO + H

d. What type of inhibition is exhibited against NAD* as a cofactor? Describe what is going on in this type of inhibition. e. Referring to the mechanism below, what step is pyrazole likely interfering with given your answers in c and d? 1. binding of NAD+ to the enzyme, LADH 2. binding of C₂H5OH to LADH 3. Oxidation of C₂H5OH (This is the reaction step) and product dissociation 4. Dissociation of NADH from LADH NAD + LADH NAD LADH + C₂H5OH NAD LADH C₂H5OH NADH LADH NAD LADH k2 NAD LADH C₂H5OH ОН NADH LADH + NADH+ LADH CH3CHO + H

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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3. Pyrozole has been proposed as a possible nontoxic inhibitor of LADH-catalyzed ethanol oxidation. Its kinetics have been studied and, in 2 separate experiments, the velocity of the reaction was measured as a function of [C₂H5OH] and of [NAD*]. The data is given in the tables below. [C₂H5OH] varies, [LADH]= 4µg/ml, [NAD+] = 350 µM, pH=7.4, 23.5C [C₂H5OH] Vo (relative Vo (relative units) 1x10-5 M pyrazole M units) No pyrazole 2.1 1.8 1.5 1.35 0.008 0.002 0.001 0.00067 [NAD+] varies, [LADH]= 4µg/ml, [C₂H5OH]= 5 mM, pH=7.4, 23.5C [NAD+] Vo (relative units) 4x10-5 M pyrazole Vo (relative units) No pyrazole 0.00011 1.7 0.000056 1.4 0.000033 1.3 0..000017 0.95 1.7 1.0 0.69 0.52 0.85 0.71 0.63 0.47 a. Plot the data as Lineweaver-Burke plots on two separate graphs. b. Determine the Km values for [C₂H5OH] and of [NAD+]. (You can't find Vmax because the exact units of Vo are not provided) c. What type of inhibition is exhibited by pyrazole against C₂H5OH is a substrate? Describe what is going on in this type of inhibition. d. What type of inhibition is exhibited against NAD+ as a cofactor? Describe what is going on in this type of inhibition. e. Referring to the mechanism below, what step is pyrazole likely interfering with given your answers in c and d? 2. binding of C₂H5OH to LADH 1. binding of NAD+ to the enzyme, LADH 3. Oxidation of C₂H5OH (This is the reaction step) and product dissociation 4. Dissociation of NADH from LADH NAD* + LADH NAD LADH + C₂H5OH + NAD LADH C₂H5OH NADH LADH k4 -4 NAD LADH + NAD LADH C₂H5OH ОН NADH LADH + NADH+LADH CH, CHO + H JL
Transcribed Image Text:3. Pyrozole has been proposed as a possible nontoxic inhibitor of LADH-catalyzed ethanol oxidation. Its kinetics have been studied and, in 2 separate experiments, the velocity of the reaction was measured as a function of [C₂H5OH] and of [NAD*]. The data is given in the tables below. [C₂H5OH] varies, [LADH]= 4µg/ml, [NAD+] = 350 µM, pH=7.4, 23.5C [C₂H5OH] Vo (relative Vo (relative units) 1x10-5 M pyrazole M units) No pyrazole 2.1 1.8 1.5 1.35 0.008 0.002 0.001 0.00067 [NAD+] varies, [LADH]= 4µg/ml, [C₂H5OH]= 5 mM, pH=7.4, 23.5C [NAD+] Vo (relative units) 4x10-5 M pyrazole Vo (relative units) No pyrazole 0.00011 1.7 0.000056 1.4 0.000033 1.3 0..000017 0.95 1.7 1.0 0.69 0.52 0.85 0.71 0.63 0.47 a. Plot the data as Lineweaver-Burke plots on two separate graphs. b. Determine the Km values for [C₂H5OH] and of [NAD+]. (You can't find Vmax because the exact units of Vo are not provided) c. What type of inhibition is exhibited by pyrazole against C₂H5OH is a substrate? Describe what is going on in this type of inhibition. d. What type of inhibition is exhibited against NAD+ as a cofactor? Describe what is going on in this type of inhibition. e. Referring to the mechanism below, what step is pyrazole likely interfering with given your answers in c and d? 2. binding of C₂H5OH to LADH 1. binding of NAD+ to the enzyme, LADH 3. Oxidation of C₂H5OH (This is the reaction step) and product dissociation 4. Dissociation of NADH from LADH NAD* + LADH NAD LADH + C₂H5OH + NAD LADH C₂H5OH NADH LADH k4 -4 NAD LADH + NAD LADH C₂H5OH ОН NADH LADH + NADH+LADH CH, CHO + H JL
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