3. Pyrozole has been proposed as a possible nontoxic inhibitor of LADH-catalyzed ethanol oxidation. Itskinetics have been studied and, in 2 separate experiments, the velocity of the reaction was measured as afunction 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 (relativeunits)Vo (relative units)1x10-5 M pyrazoleMNo pyrazole2.11.81.51.350.0080.0020.0010.00067Vo (relativeunits)No pyrazole0.000111.70.0000561.40.0000331.30..000017 0.951.71.0[NAD+] varies, [LADH]= 4µg/ml, [C₂H5OH]= 5 mM, pH=7.4, 23.5C[NAD+]Vo (relative units)4x10-5 M pyrazole0.690.520.850.710.630.47a. 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 Voare not provided)c. What type of inhibition is exhibited by pyrazole against C₂H5OH is a substrate? Describe what is going on inthis type of inhibition.2. binding of C₂2H5OH to LADHd. What type of inhibition is exhibited against NAD+ as a cofactor? Describe what is going on in this type ofinhibition.e. Referring to the mechanism below, what step is pyrazole likely interfering with given your answers in c andd?1. binding of NAD+ to the enzyme, LADH3. Oxidation of C₂H5OH (This is the reactionstep) and product dissociation4. Dissociation of NADH from LADHNAD++ LADHNAD LADH +NADk₁k_1NADH LADHC₂H5OHLADH C₂H5OH+NAD LADHk2NAD LADH C₂H5OHОНNADH LADH +NADH+ LADHCH3CHO + H

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Biochemistry

6th Edition

ISBN:9781305577206

Author:Reginald H. Garrett, Charles M. Grisham

Publisher:Reginald H. Garrett, Charles M. Grisham

Chapter27: Metabolic Integration And Organ Specialization

Section: Chapter Questions

Problem 4P

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1 a) What is meant by the ATP currency exchange ratio? Why does the oxidation of mitochondrial FADH2 generate one less ATP than oxidation of mitochondrial NADH? b) If 12 H+ are moved across the inner mitochondrial membrane by NADH oxidation, and each ATP synthesized requires 3 H+ to move through ATP synthase, why are only 3 ATP molecules produced by oxidation of each NADH?
A new drug, Proinebrium, that reduces Kcat (Ki = 2.0 uM) has been developed to treat ethylene glycol poisoning. (1) What concentration of Proinebrium is required to achieve 50% inhibition of ethylene glycol metabolism by alcohol dehydrogenase when the concentraion of ethlyene glycol in the blood is 50 uM?
What is the reaction potential for oxidation of NADH (for completely passing electrons from NADH to O2) with units? 2. What is the reaction potential for oxidation of FADH2 (from FADH2 to O2) with units? 3. Which reaction has a more negative deltaG?
A mutant version of DADH can use NADP+ as a cofactor for isopropanol oxidation. Velocity data was collected from reactions at a series of NADP+ concentrations. The following trendline was obtained for a Lineweaver-Burk plot of the data: y = 0.00007x + 0.0014 Note that the NADP+ substrate concentrations are in mM and the reaction velocity was measured in nmol/min. Calculate the Km and Vmax for DADH with this substrate. Show your work.
Based on the definition of kcat, substitute a value that can be measured and yet still represents the value associated with the original concentration of the R. What would the rate or velocity of the reaction be equal to under these circumstances? How can cells increase Vmax? What variable that we could change would directly impact Vmax? Would the value of KM be affected by the ways you determined that Vma,x could be increased? What does this indicate about KM? Thinking about how catalysts work, about the Michaelis-Menten Equation, and the definition of kcat, what specifically does the enzyme change in the reaction mechanism to increase the rate? If an enzyme follows the 2 step mechanism proposed by Michaelis-Menten, what do you know about this enzyme? Be very specific and comprehensive. Please answer very soon will give rating surely
1. A cell in an active catabolic steady state state needs to maintain A. a high cytosolic (ATP/ADP) and a high cytosolic (NADH/NAD+) ratio. B. a high cytosolic (ATP/ADP) and a low cytosolic (NADH/NAD+) ratio. C. a low cytosolic (ATP/ADP) and a low cytosolic (NADH/NAD+) ratio. D. a low cytosolic (ATP/ADP) and a high cytosolic (NADH/NAD+) ratio.
Understanding the Action of Fluoroacetate on the TCA Cycle In a tissue where the TCA cycle has been inhibited by fluoroacetate, what difference in the concentration of each TCA cycle metabolite would you expect, compared with a normal, uninhibited tissue?
Understanding the Mechanism of the -Ketoglutarate Dehydrogenase Reaction tshe first step of the -Ketoglutarate dehydrogenase reaction involves decarboxylation of the substrate and leaves a covalent TPP intermediate. Write a reasonable mechanism for this reaction.
Distinguishing the Mechanisms of Class I and Class I Aldolases Fructose bisphosphate aldolase in animal muscle is a class 1 aldolase, which forms a Schiff base intermediate between substrate (for example. fructose-1, 6-bisphosphate or dihydroxyacetone phosphate) and a lysine at the active site (see Figure I8.12). The chemical evidence for this intermediate conies from studies with aldolase and the reducing agent sodium borohydride, NaBH4. Incubation of the enzyme with dihydroxyacetone phosphate and NaBH4 inactivates the enzyme. Interestingly, no inactivation is observed if NabH4 is added to the enzyme in the absence of substrate. Write a mechanism that explains these observations and provides evidence for the formation of a Schiff base intermediate in the aldolase reaction.
A dialyzed pigeon liver extract will catalyze the conversion of acetyl-CoAto palmitate and CoASH if supplied with Mg2+, NADPH, ATP, HCO3-, andcitrate.(a) If H14CO3– is supplied, what compounds will become labeled (permanently or transiently) during the course of the reaction? In whatcompounds will 14C accumulate?(b) Explain the role of citrate in this reaction.
Does the complete oxidation of tridecanoic acid (C13:0) make more ATP than the complete oxidation of 9-tetradecenoic acid (C14:1, Δ9) inside human body? Show your calculation.
1. Consider the oxidation of the fatty acid with the common name arachidic acid. a. Draw the structure of arachidic acid. b. How many turns of the fatty acid oxidation cycle is required for the complete oxidation of arachidic acid? c. How many moles of ATP are formed from one mole of arachidic acid if the acetyl CoA produced go to the citric acid cycle and oxidative phosphorylation? Assume 1 mole of NADH is equivalent to 3 moles ATP and 1 mole FADH2 is equivalent to 2 moles of ATP. Show how you arrived at your answer

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