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Expedited isolation of natural product peptidyl-tRNA hydrolase inhibitors from a Pth1 affinity column

Department of Chemistry, University of Alabama in Huntsville, Huntsville, AL, USA

Topical Section: Enzyme Activity and Immobilization

New antibiotics and new antibiotic targets are needed to counter the development of bacterial drug resistance that threatens to return the human population to the pre-antibiotic era. Bacterial peptidyl-tRNA hydrolase (Pth1) is a promising new antibiotic target in the early stages of development. While inhibitory activity has been observed in a variety of natural products, bioactive fractionation has been a bottleneck for inhibitor isolation. To expedite the isolation of inhibitory compounds from complex mixtures, we constructed a Pth1 affinity column and used it to isolate inhibitory compounds from crude natural products. Recombinantly produced S. typhimurium Pth1 was covalently attached to a column matrix and the inhibitory activity isolated from ethanol extracts of Salvinia minima. The procedure reported here demonstrates that isolation of Pth1 inhibitory compounds from complex natural product extracts can be greatly expedited over traditional bioactive fractionation, decreasing time and expense. The approach is generally applicable to Pth1s from other bacterial species and opens an avenue to advance and accelerate inhibitor development against this promising antimicrobial target.
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References

1. Lee LA, Puhr ND, Maloney EK, et al. (1994) Increase in antimicrobial-resistant Salmonella infections in the United States, 1989-1990. J Infect Dis 170: 128-134.    

2. Glynn MK, Bopp C, Dewitt W, et al. (1998) Emergence of multidrug-resistant Salmonella enterica serotype typhimurium DT104 infections in the United States. New Engl J Med 338: 1333-1338.    

3. Centers for Disease Control and Prevention: Antibiotic/Antimicrobial Resistance. Centers for Disease Control and Prevention, 2017. Available from: https://www.cdc.gov/drugresistance/

4. Kariuki S, Gordon MA, Feasey N, et al. (2015) Antimicrobial resistance and management of invasive Salmonella disease. Vaccine 33 Suppl 3: C21-29.

5. Das G, Varshney U (2006) Peptidyl-tRNA hydrolase and its critical role in protein biosynthesis. Microbiology 152: 2191-2195.    

6. Hernandez-Sanchez J, Valadez JG, Herrera JV, et al. (1998) lambda bar minigene-mediated inhibition of protein synthesis involves accumulation of peptidyl-tRNA and starvation for tRNA. EMBO J 17: 3758-3765.

7. Cruz-Vera LR, Hernandez-Ramon E, Perez-Zamorano B, et al. (2003) The rate of peptidyl-tRNA dissociation from the ribosome during minigene expression depends on the nature of the last decoding interaction. J Biol Chem 278: 26065-26070.

8. Tenson T, Herrera JV, Kloss P, et al. (1999) Inhibition of translation and cell growth by minigene expression. J Bacteriol 181: 1617-1622.

9. Fromant M, Schmitt E, Mechulam Y, et al. (2005) Crystal structure at 1.8 A resolution and identification of active site residues of Sulfolobus solfataricus peptidyl-tRNA hydrolase. Biochemistry 44: 4294-4301.

10. Powers R, Mirkovic N, Goldsmith-Fischman S, et al. (2005) Solution structure of Archaeglobus fulgidis peptidyl-tRNA hydrolase (Pth2) provides evidence for an extensive conserved family of Pth2 enzymes in archea, bacteria, and eukaryotes. Protein Sci 14: 2849-2861.    

11. Jan Y, Matter M, Pai JT, et al. (2004) A mitochondrial protein, Bit1, mediates apoptosis regulated by integrins and Groucho/TLE corepressors. Cell 116: 751-762.    

12. Rosas-Sandoval G, Ambrogelly A, Rinehart J, et al. (2002) Orthologs of a novel archaeal and of the bacterial peptidyl-tRNA hydrolase are nonessential in yeast. Proc Natl Acad Scie U S A 99: 16707-16712.    

13. Ito K, Murakami R, Mochizuki M, et al. (2012) Structural basis for the substrate recognition and catalysis of peptidyl-tRNA hydrolase. Nucleic Acids Res 40: 10521-10531.    

14. Giorgi L, Bontems F, Fromant M, et al. (2011) RNA-binding site of Escherichia coli peptidyl-tRNA hydrolase. J Biol Chem 286: 39585-39594.    

15. Hames MC, McFeeters H, Holloway WB, et al. (2013) Small molecule binding, docking, and characterization of the interaction between Pth1 and peptidyl-tRNA. Int J Mol Sci 14: 22741-22752.    

16. McFeeters H, Gilbert MJ, Thompson RM, et al. (2012) Inhibition of essential bacterial peptidyl-tRNA hydrolase activity by tropical plant extracts. Nat Prod Commun 7: 1107-1110.

17. Kaushik S, Singh N, Yamini S, et al. (2013) The mode of inhibitor binding to peptidyl-tRNA hydrolase: binding studies and structure determination of unbound and bound peptidyl-tRNA hydrolase from Acinetobacter baumannii. PloS One 8: e67547.    

18. Giorgi L, Plateau P, O'Mahony G, et al. (2011) NMR-based substrate analog docking to Escherichia coli peptidyl-tRNA hydrolase. J Mol Biol 412: 619-633.    

19. Ferguson PP, Holloway WB, Setzer WN, et al. (2016) Small Molecule Docking Supports Broad and Narrow Spectrum Potential for the Inhibition of the Novel Antibiotic Target Bacterial Pth1. Antibiotics 5: 16.    

20. Kabra A, Shahid S, Pal RK, et al. (2017) Unraveling the stereochemical and dynamic aspects of the catalytic site of bacterial peptidyl-tRNA hydrolase. RNA 23: 202-216.    

21. Goodall JJ, Chen GJ, Page MG (2004) Essential role of histidine 20 in the catalytic mechanism of Escherichia coli peptidyl-tRNA hydrolase. Biochemistry 43: 4583-4591.    

22. Fromant M, Plateau P, Schmitt E, et al. (1999) Receptor site for the 5'-phosphate of elongator tRNAs governs substrate selection by peptidyl-tRNA hydrolase. Biochemistry 38: 4982-4987.    

23. Taylor-Creel K, Hames MC, Holloway WB, et al. (2014) Expression, purification, and solubility optimization of peptidyl-tRNA hydrolase 1 from Bacillus cereus. Protein Expr Purif 95: 259-264.    

24. Holloway WB, McFeeters H, Powell AM, et al. (2015) A Highly Adaptable Method for Quantification of Peptidyl-tRNA Hydrolase Activity. J Anal Bioanal Tech 6: 244.

25. McFeeters H, McFeeters RL (2014) Current Methods for Analysis of Enzymatic Peptidyl-tRNA Hydrolysis. J Anal Bioanal Tech 5: 215.

26. El-Elimat T, Raja HA, Day CS, et al. (2017) alpha-Pyrone derivatives, tetra/hexahydroxanthones, and cyclodepsipeptides from two freshwater fungi. Bioorg Med Chem 25: 795-804.    

27. Harris SM, McFeeters H, Ogungbe IV, et al. (2011) Peptidyl-tRNA hydrolase screening combined with molecular docking reveals the antibiotic potential of Syzygium johnsonii bark extract. Nat Prod Commun 6: 1421-1424.

28. Bonin PD, Erickson LA (2002) Development of a fluorescence polarization assay for peptidyl-tRNA hydrolase. Anal Biochem 306: 8-16.    

Copyright Info: © 2017, Robert L. McFeeters, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

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