Cloning, expression, purification and kinetics of Trehalose-6-phosphate phosphatase of filarial parasite Brugia malayi
IR@CDRI: CSIR-Central Drug Research Institute, Lucknow
View Archive Info| Field | Value | |
| Creator |
Susheela, Kushwaha
Singh, P K Rana, A K Bhattacharya-Misra, Shailja |
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| Date |
2011-09-30T10:41:27Z
2011-09-30T10:41:27Z 2011 |
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| Identifier |
Acta Tropica, (2011), 119(2-3), 151-159
http://hdl.handle.net/123456789/724 |
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| Description |
The pleiotropic functions of disaccharide trehalose in the biology of nematodes and its absence from mammalian cells suggest that its biosynthesis may provide a useful target for developing novel nematicidal drugs. The trehalose-6-phosphate phosphatase (TPP), one of the enzymes of trehalose metabolism has not been characterized so far in nematodes except the free living nematode Caenorhabditis elegans where it’s silencing results in to lethal outcomes. This prompted us to clone and characterize B. malayi TPP in order to discover novel antifilarial drug target. The recombinant protein (Bm-TPP) was purified with apparent homogeneity on a metal ion column and it was found to possess high phosphatase activity with robust specificity for the substrate trehalose-6-phosphate. Bm-TPP was found to be a member of the HAD-like hydrolase super family II based on the conserved motifs required for catalytic reaction. The Km for substrate trehalose-6-Phosphate was around 0.42 mM with pH optimum ~7.0 and the enzyme showed an almost absolute requirement for Mg2+ as a metal ion. Bm-TPP was expressed in all the life-stages of B. malayi. In the absence of an effective macrofilaricidal agent and validated antifilarial drug target, Bm-TPP bodes well as a rational drug target against lymphatic filariasis.
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630924 bytes
application/pdf |
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| Language |
en
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| Relation |
CDRI COMMUNICATION NO. 8072
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| Subject |
Brugia malayi
lymphatic filariasis antifilarial drug target trehalose-6-phosphate phosphatase |
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| Title |
Cloning, expression, purification and kinetics of Trehalose-6-phosphate phosphatase of filarial parasite Brugia malayi
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| Type |
Article
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