Abstract
Aminoacyl-tRNA protein transferases post-translationally conjugate an amino acid from an aminoacyl-tRNA onto the N-terminus of a target polypeptide. The eubacterial aminoacyl-tRNA protein transferase, L/F transferase, utilizes both leucyl-tRNALeu and phenylalanyl-tRNAPhe as substrates. X-ray crystal structures with substrate analogues, the minimal substrate phenylalanyl adenosine (rA-Phe) and inhibitor puromycin, have been used to characterize tRNA recognition by L/F transferase. However analyses of these two X-ray crystal structures reveal significant differences in binding. Through structural analyses, mutagenesis, and enzymatic activity assays, we rationalize and demonstrate that the substrate analogues bind to L/F transferase with similar binding affinities using a series of different interactions by the various chemical groups of the analogues. Our data also demonstrates that enlarging the hydrophobic pocket of L/F transferase selectively enhances puromycin inhibition and may aid in the development of improved inhibitors for this class of enzymes.
Keywords: Aminoacyl-tRNA protein transferase, L/F transferase, N-end rule, puromycin, quantitative mass spectrometry.
Related Books
Applied Biomathematics for Nucleic Acid Chemistry and Protein Folding: Quantitative Simulations
Industrial Applications of Soil Microbes
Molecular and Physiological Insights into Plant Stress Tolerance and Applications in Agriculture
Systems Biology, Bioinformatics and Livestock Science
Advances in Legume Research: Physiological Responses and Genetic Improvement for Stress Resistance
Alternative Remedies and Natural Products for Cancer Therapy: An Integrative Approach
Future Farming: Advancing Agriculture with Artificial Intelligence
The Drone Honey Bee
Fungal Lipid Biochemistry
Steroids and their Medicinal Potential
28