mm Page S-211 pis vast . EQA
chapter
Amino Acid Oxidation
and the Production
of Urea
1, Products of Amino Acid Transamination Name and draw the structure of the a-keto acid
resulting when each ofthe following amino acids undergoes transamination with a-ketoglutarate
(a) aspartate, (b) slutamate, (e) alanine, (4) phenylalanine.
Answer
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2, Measurement of Alanine Aminotransferase Activity The activity (reaction rate) of alanine
aminotransferase is usually measured by including an excess of pure lactate deliydrogenase and NADH
{in the reaction system. The rate of alanine disappearance is equal to the rate of NADH disappearance
‘measured spectrophotometricaly. Explain how this assay works.
Answer The measurement of the activity of alanine aminotransferase by measurement of the
reaction of its product with lactate dehydrogenase is an example of a “coupled” assay. The prod
uct ofthe transamination (pyruvate) is rapidly consumed in the subsequent “indicator reaction,”
catalyzed by an excess of lactate dehydrogenase. The dehydrogenase uses the cofactor NADH,
the disappearance of which is conveniently measured by observing the rate of decrease in
[NADH absorption at 340 nm. Thus, the rate of disappearance of NADH is a measure of the rate
of the aminotransferase reaction, ‘(NADH and lactate dehydrogenase are added in excess.
‘3, Alanine and Glutamine in the Blood Normal human blood plasma contains all the amino acids re
{quired for the synthesis of body proteins, but notin equal concentrations. Alanine and glutamine are
present in much higher concentrations than any other amino acids. Suggest why.
‘Answer Musele tissue ean convert amino acids to thelr Keto acids plus ammonia, then oxidize
‘the keto acids to produce ATP for muscle contraction. However, urea cannot be formed in
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$-212 Chapter 18 Amin Acié Oxidation ané the Production of Urea
‘muscle. Alanine and glutamine transport amino groups in the bloodstream to the liver (see
Fig. 18-2) from muscle and other nonhepatic tissues. In muscle, amino groups from all amino
acids are transferred to pyruvate or glutamate to form alanine or glutamine, and these latter
amino acids are transported to the liver.
4, Distribution of Amino Nitrogen If your diet src in alanine but deficient in aspartate, will you
show signs of aspartate deficiency? Explain.
Answer No; aspartate s realy formed by the transfer of the amino group of alanine to ox-
aloacetate, Celular levels of aminotransferases are sufficient to provide all of the amino acids
in this fashion, if the a-keto acids are available.
5, Lactate versus Alanine as Metabolic Fuel: The Cost of Nitrogen Removal The three carbons in
lactate and alanine have identical oxidation states, and animals can use either carbon source as a meta-
Dolie fuel. Compare the net ATP yield (moles of ATP per mole of substrate) for the complete oxidation
(to CO, and H,0) of lactate versus alanine when the cost of nitrogen excretion as urea is included,
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Answer Lactate and alanine are converted to pyruvate by their respective dehydrogenases,
lactate dehydrogenase and alanine dehydrogenase, producing pyruvate and NADH + I" and,
‘in the case of alanine, NH. Complete oxidation of T mol of pyruvate to CO, and H,0 produces
12,6 mol of ATP via the citric acid cycle and oxidative phosphorylation (see Table 16-1). In
addition, the NADH from each dehydrogenase reaction produces 2.5 mol of ATP per mole of
[NADH reoxidized. Thus oxidation produces 16 mol of ATP per mole of lactate. Urea formation
uses the equivalent of 4 mol of ATP per mole of urea formed (Fig. 18-10), or 2 mol of ATP per
‘mol of NH. Subtracting this value from the energy yield of alanine results in 13 mol of ATP.
per mole of alanine oxidized.
6. Ammonia Toxicity Resulting from an Arginine-Deficient Diet In a study conducted some years
‘ago, cats were fasted overnight then given a single meal complete in all amino aeids except arginine
Within 2 hours, blood ammonia levels increased from a normal level of 18 pa/L to 140 pg/L, and the cats
showed the clinical symptoms of ammonia toxicity. A control group fed a complete amino acid diet or
fn amino acid dlet in which arginine was replaced by ornithine showed no unusual clinical symptoms
(a) What was the role of fasting in the experiment?
Cb). What cansed the arumonia levels to rise in the experimental group? Why did the absence of arg
rine lead to ammonia toxicity? Is arginine an essential amino acid in cats? Why or wity not?
(e)_ Why can omithine be substituted for arginine?
Answer
(a) Festing resulted in lowering of blood glucose levels. Subsequent feeding of an arginine-
free diet led to a rapid catabolism ofall the ingested amino acl, especially the gluco-
‘genic ones. This catabolism was exacerbated by the lack of an essential amino acid,
Which prevented protein synthesis,
(b) Oxidative deamination of amino acids caused the elevation of ammonia levels. In ada
tion, the lack of arginine (an intermediate in the urea cycle) slovred the conversion of
ammonia to urea. Arginine (or ornithine) synthesis inthe cat is not sufficient to meet
the needs imposed by the stress of this experiment, suggesting that arginine is an essen-
tial amino acid
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ocketoglutarate + NH + NADH + Ht NH] + 2ATP + 1,0 + CO, —> carbamoyl phosphate + 24DP + P, + SH* Carbamoyl phosphate + ornithine — citrulline + P, + H* Chtruline + aspartate + ATP —* argininoeuccinate + AMP + PP, +H" ‘Argininosuceinate —> arginine + fumarate Fumarate + H,O —> malate Malate + NAD* — oxaloacetate + NADH + H* Oxaloacetate + glutamate —> aspartate + a-ketoghitarate Arginine + H,0 —> urea + ornithine ‘The sum of these resetions is 2 Glutamate + CO, + 4H,0 + 2NAD* + 8ATP —> 2erketoglutarate + 2NADH + 7H" + urea + 2ADP + AMP + PP) + 22, @ "Three additional reactions need to be considered: AMP + ATP—> 2apP @ (+ 8H” + 2NADH + 6ADP + GP, —» 2NAD* + GATP + 81,0 @ HO + PP, 2P, + HY @ ‘Sumaning the last four equations: 2 Glutamate + CO, + 0, +2ADP + 2P,—> 2 a-ketoglutarate + urea + 3H,0 + 2ATP 8, Transamination and the Urea Cyele Aspartate aminotransferase has the highest activity of all the ‘mammalian liver eminotransferases, Why? Answer The second amino group introduced into urea is transferred from aspartate. This amino seid is generated in large quantities by transamination between oxaloacetate and gluta- mate (and many other amino acids), catalyzed by aspartate aminotransferase. Approximately ‘one half ofall the amino groups excreted as urea must pass through the aspartate aminotrans- {orase reaction, and liver contains higher levels ofthis aminotransferase than of any’ other. [Bj ® Te Case against the Higuld Protein Diet A weightreducing det healypromated some years ‘ago required the daily intake of “liquid protein” (soup of hydrolyzed gelatin), water, and an assortment of vitamins. All other food and drink were to be avoided. People on this diet typically lost 10 to 14 Ib in ‘the frst week (@) Opponents argued that the weightloss was almost entirely due to water loss and would be regained very soon after a normal diet was resumed. What isthe biochemical basis for this argument? Cb). Annumber of people on this diet died. What are sorte of the dangers inherent in the diet and how can they lead to death? Answer (a) A person on a diet consisting only of protein must use amino aeids as the prineipal source of metabolic fuel. Because the catabolism of amino acids requires the removal of hp CONFIRMING PAGES —Vous aimerez peut-être aussi
