Protein Engineering Multiple Choice Questions on “Applications – Engineering Tyrosyl-tRNA Synthetase”.
1. Which of the following enzyme catalyzes the charging of tRNATyr with tyrosine?
A. Tyrosine synthetase
B. RNA activating enzyme
C. Tyrosyl-tRNA synthase
D. Tyrosyl-tRNA synthetase
Answer: D
Clarification: Tyrosyl-tRNA synthetase catalyzes the charging of tRNATyr with tyrosine. It is a two-stage reaction in which the tyrosine is first activated at its carboxyl group with ATP to form tyrosyl adenylate. Tyrosine synthetase, RNA activating enzyme, or Tyrosyl-tRNA synthase do not catalyze the charging of tRNATyr with tyrosine.
2. The C-terminal domain of tyrosyl-tRNA synthetase contains major binding determinants for the tRNA.
A. False
B. True
Answer: B
Clarification: The above statement is true. The C-terminal domain of tyrosyl-tRNA synthetase contains major binding determinants for the tRNA. The C-terminal domain was excised at the level of the cloned gene and the kinetics of the N-terminal domain was assayed. The kinetics of the tyrosine activation were identical for the wild type and the truncated enzyme, but the truncated enzyme did not charge or bind tRNA.
3. Which of the following shaped structure is formed by the tRNATyr ?
A. J-shaped
B. O-shaped
C. I-shaped
D. L-shaped
Answer: D
Clarification: tRNATyr folds into an L-shaped structure, with the receptor stem and the TψC stem forming one arm, and the anticodon stem and extra loop the other arm. tRNATyr does not form J-shaped, O-shaped, or I-shaped.
4. In tRNA charging, the Km (ATP) is unaffected in the heterodimer, but the Kcat is one half that of wild type enzyme.
A. False
B. True
Answer: B
Clarification: The above statement is true. The C-terminal domain of tyrosyl-tRNA synthetase contains major binding determinants for the tRNA. Thus, the heterodimer appears to be a fully refolded, active molecule, in which one subunit can complement a lesion on the other subunit.
5. Which of the following amino acids can be used in place of lysine, arginine, and histidine to remove the electrostatic charge but retain the hydrophilic nature of the side chain?
A. Aspartic acid and glutamic acid
B. Aspartic acid and glutamine
C. Asparagine and glutamic acid
D. Asparagine and glutamine
Answer: D
Clarification: Asparagine and glutamine can be used in place of lysine, arginine, and histidine to remove the electrostatic charge but retain the hydrophilic nature of the side chain. Aspartic acid and glutamic acid cannot be used in place of lysine, arginine, and histidine to remove the electrostatic charge and retain the hydrophilic nature of the side chain. Asparagine and glutamine contain neutral but polar side chains.
6. The charging of tRNATyr with tyrosine is a single-stage reaction.
A. True
B. False
Answer: B
Clarification: The above statement is false. The charging of tRNATyr with tyrosine is a two-stage reaction. The tyrosine is first activated at its carboxyl group with ATP to form tyrosyl adenylate. Subsequently, the adenylate is attacked by the 2’ or 3’ hydroxyl of the tRNA to give tyrosyl-tRNATyr.
7. Tyrosine is activated at an amino group with ATP to form tyrosyl adenylate.
A. True
B. False
