scholarly journals Characterization and structure determination of prolyl‐tRNA synthetase from Pseudomonas aeruginosa and development as a screening platform

2019 ◽  
Vol 28 (4) ◽  
pp. 727-737 ◽  
Author(s):  
Noah Pena ◽  
David M. Dranow ◽  
Yanmei Hu ◽  
Yaritza Escamilla ◽  
James M. Bullard
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Structure Determination ◽  
Trna Synthetase ◽  
Screening Platform

scholarly journals Glutaminyl‐tRNA Synthetase from Pseudomonas aeruginosa : Characterization, structure, and development as a screening platform

2019 ◽  
Vol 29 (4) ◽  
pp. 905-918
Author(s):  
Yaritza Escamilla ◽  
Casey A. Hughes ◽  
Jan Abendroth ◽  
David M. Dranow ◽  
Samantha Balboa ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Trna Synthetase ◽  
Screening Platform

scholarly journals Identification and Characterization of Chemical Compounds that Inhibit Leucyl-tRNA Synthetase from Pseudomonas aeruginosa

2020 ◽  
Vol 17 (1) ◽  
pp. 119-130 ◽  
Author(s):  
Regina Zamacona ◽  
Pamela N. Chavero ◽  
Eduardo Medellin ◽  
Yanmei Hu ◽  
Casey A. Hughes ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Human Cell ◽  
Bacterial Infections ◽  
Chemical Compounds ◽  
Human Cell Line ◽  
Trna Synthetase ◽  
Multi Drug Resistance ◽  
Bacterial Strains ◽  
Gene Encoding ◽  
Screening Platform

Background: Pseudomonas aeruginosa is an opportunistic multi-drug resistance pathogen implicated as the causative agent in a high-percentage of nosocomial and community acquired bacterial infections. The gene encoding leucyl-tRNA synthetase (LeuRS) from P. aeruginosa was overexpressed in Escherichia coli and the resulting protein was characterized. Methods: LeuRS was kinetically evaluated and the KM values for interactions with leucine, ATP and tRNA were 6.5, 330, and 3.0 μM, respectively. LeuRS was developed into a screening platform using scintillation proximity assay (SPA) technology and used to screen over 2000 synthetic and natural chemical compounds. Results: The initial screen resulted in the identification of two inhibitory compounds, BT03C09 and BT03E07. IC50s against LeuRS observed for BT03C09 and BT03E07 were 23 and 15 μM, respectively. The minimum inhibitory concentrations (MIC) were determined against nine clinically relevant bacterial strains. In time-kill kinetic analysis, BT03C09 was observed to inhibit bacterial growth in a bacteriostatic manner, while BT03E07 acted as a bactericidal agent. Neither compound competed with leucine or ATP for binding LeuRS. Limited inhibition was observed in aminoacylation assays with the human mitochondrial form of LeuRS, however when tested in cultures of human cell line, BT03C09 was toxic at all concentration whereas BT03E07 only showed toxic effects at elevated concentrations. Conclusion: Two compounds were identified as inhibitors of LeuRS in a screen of over 2000 natural and synthetic compounds. After characterization one compound (BT03E07) exhibited broad spectrum antibacterial activity while maintaining low toxicity against human mitochondrial LeuRS as well as against human cell cultures.

Ab initio structure determination of cytochrome c6 by combined reciprocal space-real space approach

2003 ◽  
Vol 218 (1) ◽  
Author(s):  
K. Chowdhury ◽  
S. Ghosh ◽  
M. Mukherjee
Keyword(s):  
Cytochrome C ◽  
Ab Initio ◽  
Structure Determination ◽  
Direct Method ◽  
Real Space ◽  
Reciprocal Space ◽  
Cytochrome C6 ◽  
Ab Initio Structure ◽  
Space Approach

AbstractThe direct method program SAYTAN has been applied successfully to redetermine the structure of cytochrome c

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