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| The Establishment of an Anti-Trypanosoma Drug Screening System with Leucyl-tRNA Synthetase as an Inhibition Target |
| GAO Guang-wei,YAO Ying,DING Da-zhong,YE Long,ZHOU Hu-chen,LI Da-wei |
| The Institute of Chemical Biology, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China |
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Abstract Trypanosoma is a human parasite severely affecting poor tropical areas. However, current frontline drugs for Trypanosoma treatment have severe side-effects with decreased effectiveness. Based on the fact that aminoacyl-tRNA synthetase is a bona fide drug target for several microorganisms, including bacteria and fungi, it is plausible that it may also be effective target of Trypanosoma. In this report, the Trypanosoma brucei Leucyl-tRNA synthetase (tbLeuRS) was cloned , expressed and purified to develop an in vitro enzymatic assay system. The assay conditions were further optimized for the effective screening of tbLeuRS inhibitors thus establishing an anti-Trypanosoma drug screening system targeting tbLeuRS. The results indicated that this system can be employed for the effective screening of anti-Trypanosoma drugs with satisfactory specificity. In addition, this system can also be used for compound optimization, as well as IC50 testing. Using this system a series of compounds are identified that are effective Trypanosoma inhibitors without toxicity to human cells. Therefore, targeting tbLeuRS may represent a new venue for the development of anti-Trypanosoma drugs.
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Received: 09 July 2009
Published: 21 December 2009
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| ［1］ |
Fevre E M, Coleman P G, Odiit M, et al.The origins of a new Trypanosoma brucei rhodesiense sleeping sickness outbreak in eastern Uganda. Lancet, 2001, 358(9282): 625~628
|
|
|
| ［2］ |
MacLeod A, Tait ATurner C M. The population genetics of Trypanosoma brucei and the origin of human infectivity. Philos Trans R Soc Lond B Biol Sci, 2001, 356(1411): 1035~1044
|
|
|
| ［3］ |
Thuita J K, Kagira J M, Mwangangi D, et al.Trypanosoma brucei rhodesiense transmitted by a single tsetse fly bite in vervet monkeys as a model of human African trypanosomiasis. PLoS Negl Trop Dis, 2008, 2(5): e238
|
|
|
| ［4］ |
Schultzberg M, Ambatsis M, Samuelsson E B, et al. Spread of Trypanosoma brucei to the nervous system: early attack on circumventricular organs and sensory ganglia. J Neurosci Res, 1988, 21(1): 56~61
|
|
|
| ［5］ |
Woods A, Sherwin T, Sasse R, et al. Definition of individual components within the cytoskeleton of Trypanosoma brucei by a library of monoclonal antibodies. J Cell Sci, 1989, 93(Pt 3): 491~500
|
|
|
| ［6］ |
Lee S W, Cho B H, Park S G, et al. AminoacyltRNA synthetase complexes: beyond translation. J Cell Sci, 2004, 117(Pt 17): 3725~3734
|
|
|
| ［7］ |
Pohlmann J,BrotzOesterhelt H. New aminoacyltRNA synthetase inhibitors as antibacterial agents. Curr Drug Targets Infect Disord, 2004, 4(4): 261~272
|
|
|
| ［8］ |
Rock F L, Mao W, Yaremchuk A, et al. An antifungal agent inhibits an aminoacyltRNA synthetase by trapping tRNA in the editing site. Science, 2007, 316(5832): 1759~1761
|
|
|
| ［9］ |
Hausmann C DIbba M. AminoacyltRNA synthetase complexes: molecular multitasking revealed. FEMS Microbiol Rev, 2008, 32(4): 705~721
|
|
|
| ［10］ |
Green L S, Bullard J M, Ribble W, et al. Inhibition of methionyltRNA synthetase by REP8839 and effects of resistance mutations on enzyme activity. Antimicrob Agents Chemother, 2009, 53(1): 86~94
|
|
|
| ［11］ |
Tandon M, Coffen D L, Gallant P, et al. Potent and selective inhibitors of bacterial methionyl tRNA synthetase derived from an oxazolonedipeptide scaffold. Bioorg Med Chem Lett, 2004, 14(8): 1909~1911
|
|
|
| ［12］ |
Yao P, Zhou X L, He R, et al. Unique residues crucial for optimal editing in yeast cytoplasmic LeucyltRNA synthetase are revealed by using a novel knockout yeast strain. J Biol Chem, 2008, 283(33): 22591~22600
|
|
|
| ［13］ |
Brown M J, Mensah L M, Doyle M L, et al. Rational design of femtomolar inhibitors of isoleucyl tRNA synthetase from a binding model for pseudomonic acidA. Biochemistry, 2000, 39(20): 6003~6011
|
|
|
| ［14］ |
Roth A, Eriani G, Dirheimer G, et al. Kinetic properties of pure overproduced Bacillus subtilis phenylalanyltRNA synthetase do not favour its in vivo inhibition by ochratoxin A. FEBS Lett, 1993, 326(13): 87~91
|
|
|
| ［15］ |
Lue S WKelley S O. An aminoacyltRNA synthetase with a defunct editing site. Biochemistry, 2005, 44(8): 3010~3016
|
|
|
| ［16］ |
Boniecki M T, Vu M T, Betha A K, et al. CP1dependent partitioning of pretransfer and posttransfer editing in leucyltRNA synthetase. Proc Natl Acad Sci U S A, 2008, 105(49): 19223~19228
|
|
|
| ［17］ |
Puglisi J D, Putz J, Florentz C, et al. Influence of tRNA tertiary structure and stability on aminoacylation by yeast aspartyltRNA synthetase. Nucleic Acids Res, 1993, 21(1): 41~49
|
|
|
| ［18］ |
Gehrig SEfferth T. Development of drug resistance in Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense. Treatment of human African trypanosomiasis with natural products. Int J Mol Med, 2008, 22(4): 411~419
|
|
|
| ［19］ |
Hurdle J G, O'Neill A JChopra I. Prospects for aminoacyltRNA synthetase inhibitors as new antimicrobial agents. Antimicrob Agents Chemother, 2005, 49(12): 4821~4833
|
|
|
| ［20］ |
Rho S B, Lincecum T L,Martinis S A. An inserted region of leucyltRNA synthetase plays a critical role in group I intron splicing. EMBO J, 2002, 21(24): 6874~6881
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