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Crystallization of uracil phosphoribosyltransferase (MtUPRT) from Mycobacterium tuberculosis

Ghode, Pramila Baban and Sivaraman , Jayaraman and Chacko , Jobichen and Bifani, Juan Pablo (2015) Crystallization of uracil phosphoribosyltransferase (MtUPRT) from Mycobacterium tuberculosis. Biochemical and biophysical research communications : BBRC, 467 (3). pp. 577-582. ISSN 0006291X

Abstract

Exploring new drug targets in parallel to designing strategies for rational use of existing drugs would greatly aid the Tuberculosis (TB) drug development program. The key enzymes involved in the essential metabolic and regulatory pathways are usually sought for in the pursuit of potential drug targets. Likewise, uracil phosphoribosyltransferase (UPRT) is a key enzyme in the synthesis uridine 5’-monophosphate (UMP), the precursor of the pyrimidine nucleotides. It has been recently shown to be the probable target of 5-fluorouracil in Mycobacterium tuberculosis (Mtb). Here we report the purification, characterization and crystallization of the full length UPRT from Mtb (MtUPRT) encoded by the gene upp (Rv3309c). The MtUPRT was overexpressed in BL21 (DE3) E.coli expression system followed by three step chromatographic purification procedures. The purified MtUPRT was concentrated to 8mg/ml; single crystals were obtained using the sitting drop vapour diffusion method. The crystals were diffracted to 3.0 Å resolution and belonged to the space group P32 with unit cell parameters a = b = 118.09, c = 77.88 Å and four monomers in the asymmetric unit. Understanding the three dimensional structure of this essential enzyme will greatly help in screening of appropriate inhibitors of MtUPRT and thus assist in TB drug development.

Item Type: Article
Keywords: Crystals, Uracil phosphoribosyltransferase (UPRT), Mycobacterium tuberculosis (Mtb), upp (Rv3309c)
Date Deposited: 12 Oct 2016 00:45
Last Modified: 12 Oct 2016 00:45
URI: https://oak.novartis.com/id/eprint/26379

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