Assessment of methylthioadenosine/S-adenosylhomocysteine nucleosidases of Borrelia burgdorferi as targets for novel antimicrobials using a novel high-throughput method.

 J Antimicrob Chemother. 2009 Jun;63 6:1163-72. Epub 2009 Apr 17. Assessment of 
methylthioadenosine/S-adenosylhomocysteine nucleosidases of Borrelia burgdorferi as targets for 
novel antimicrobials using a novel high-throughput method. Cornell KA, Primus S, Martinez JA, 
Parveen N.
Department of Chemistry and Biochemistry, Boise State University, IA 83725-1520, USA.
Abstract
BACKGROUND: Lyme disease is the most prevalent tick-borne disease in the USA with the 
highest number of cases (27 444 patients) reported by CDC in the year 2007, representing an 
unprecedented 37% increase from the previous year. The haematogenous spread of Borrelia
burgdorferi to various tissues results in multisystemic disease affecting the heart, joints, skin, 
musculoskeletal and nervous system of the patients. OBJECTIVES: Although Lyme disease can 
be effectively treated with doxycycline, amoxicillin and cefuroxime axetil, discovery of novel 
drugs will benefit the patients intolerant to these drugs and potentially those suffering from 
chronic Lyme disease that is refractory to these agents and to macrolides. In this study, we 
have explored 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase as a drug target for 
B. burgdorferi, which uniquely possesses three genes expressing homologous enzymes with two 
of these proteins apparently exported. METHODS: The recombinant B. burgdorferi Bgp and Pfs
proteins were first used for the kinetic analysis of enzymatic activity with both substrates and 
with four inhibitors. We then determined the antispirochaetal activity of these compounds using 
a novel technique. The method involved detection of the live-dead B. burgdorferi by fluorometric
analysis after staining with a fluorescent nucleic acids stain mixture containing Hoechst 33342 
and Sytox Green. RESULTS: Our results indicate that this method can be used for 
high-throughput screening of novel antimicrobials against bacteria. The inhibitors formycin A 
and 5'-p-nitrophenythioadenosine particularly affected B. burgdorferi adversely on prolonged 
treatment. CONCLUSIONS: On the basis of our analysis, we expect that structure-based 
modification of the inhibitors can be employed to develop highly effective novel antibiotics 
against Lyme spirochaetes




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