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Spring 2014 Name(s): Dalen Kuang (411741), Christopher Mejias (413899), Alex Jin (413433)

Bio4820 Problem Set 3: Ch 20-21: Nitrogen/Amino Acids


Due: Monday, March 3, 2014 Instructions: Up to three students may work together and turn in a single paper. Please keep answers as concise and legible as possible. All answers (except question 2) must be TYPED. (30 points total) 1. A two-year-old child was taken to the hospital. His mother said that he vomited frequently, especially after feedings. The childs weight and physical development were below normal. A urine sample treated with ferric chloride (FeCl3) gave a green color characteristic of the presence of phenylpyruvate. Quantitative analysis of urine samples gave the results shown in the table. (6 points)

a) Which enzyme might be deficient in this child? (2 points) Phenylalanine hydroxylase b) Propose a treatment in no more than 1 sentence (2 points). Diet low in phenylalanine, and/or oral administration of tetrahydroblopterin c) Tyrosine is normally a nonessential amino acid, but individuals with the deficient enzyme from part a) require tyrosine in their diet for normal growth. Please explain why in no more than 1 sentence. (2 points) Phenylalanine hydroxylase is necessary for the synthesis of Tyr from Phe.

2. When [2-14C,15N] glutamate undergoes oxidative degradation in the liver of a rat, will any of these isotopes be found in the atoms of urea? If so, draw the structure of urea and indicate which atoms will be labeled. (4 points)

Spring 2014 3. In a study conducted some years ago, cats were fasted overnight then given a single meal complete in all amino acids except arginine. Within 2 hours, blood ammonia levels increased from a normal level of 18 g/L to 140 g/L, and the cats showed the clinical symptoms of ammonia toxicity. A control group fed a complete amino acid diet or an amino acid diet in which arginine was replaced by ornithine showed no unusual clinical symptoms. (8 points) a) In no more than 2 sentences, explain the role of fasting in this experiment. (4 points) Fasting reduces the plasma levels of Arg before the experiment is conducted. Only the control group is supposed to have a dietary supply of Arg.

b) What caused the ammonia levels to rise in the experimental group? Why did the absence of arginine lead to ammonia toxicity? Please write no more than 2 sentences. (2 points) In cats, Arg is an essential amino acid, so they must obtain Arg from the diet. Arg is required for the urea cycle to occur, so deficiency prevents ammonia from being converted to urea and being excreted.

c) In no more than 1 sentence, explain why ornithine can be substituted for arginine. (2 point) Ornithine can be used to synthesize Arg. 4. Covalent modification (i.e. adenylation and deadenylation) is one way glutamine synthetase is regulated. (6 points) a) What are the respective effects of -ketoglutarate and glutamine on adenylation of glutamine synthetase? Please answer in no more than 1 sentence. (2 points) -ketoglutarate activates uridylyltransferase, which promotes de-adenylylation of GS, stimulating its activity, while glutamine has the opposite effect. b) In no more than 2 sentences, explain how this relates to nitrogen levels. (4 points) High -ketoglutarate levels imply low nitrogen levels. High glutamine levels imply high nitrogen levels. Glutamine synthesis is needed when nitrogen levels are low.

5. The active ingredient in the popular herbicide Roundup is N-(phosphonomethyl) glycine, commonly known as glyphosate. Glyphosate inhibits the production of chorismate, an intermediate in the synthesis of aromatic amino acids. Why is glyphosate lethal for plants, but safe to use around animals? Please write no more than 4 sentences. (6 pts) Humans lack the pathway for aromatic amino acid synthesis. Plants have a pathway for aromatic amino acid synthesis, which involves the conversion of skikimate into chorismate using EPSPS. Glyphosate inhibits EPSPS by binding to the active site.

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