Structure of the trehalose-6-phosphate phosphatase from Brugia malayi reveals key design principles for anthelmintic drugs.

Autor: Farelli JD; Department of Chemistry, Boston University, Boston, Massachusetts, United States of America., Galvin BD; New England Biolabs, Division of Parasitology, Ipswich, Massachusetts, United States of America., Li Z; New England Biolabs, Division of Parasitology, Ipswich, Massachusetts, United States of America., Liu C; Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, New Mexico, United States of America., Aono M; Department of Chemistry, Boston University, Boston, Massachusetts, United States of America., Garland M; Department of Chemistry, Boston University, Boston, Massachusetts, United States of America., Hallett OE; Department of Chemistry, Boston University, Boston, Massachusetts, United States of America., Causey TB; New England Biolabs, Division of Parasitology, Ipswich, Massachusetts, United States of America., Ali-Reynolds A; New England Biolabs, Division of Parasitology, Ipswich, Massachusetts, United States of America., Saltzberg DJ; Department of Chemistry, Boston University, Boston, Massachusetts, United States of America., Carlow CK; New England Biolabs, Division of Parasitology, Ipswich, Massachusetts, United States of America., Dunaway-Mariano D; Department of Chemistry and Chemical Biology, University of New Mexico, Albuquerque, New Mexico, United States of America., Allen KN; Department of Chemistry, Boston University, Boston, Massachusetts, United States of America.
Jazyk: angličtina
Zdroj: PLoS pathogens [PLoS Pathog] 2014 Jul 03; Vol. 10 (7), pp. e1004245. Date of Electronic Publication: 2014 Jul 03 (Print Publication: 2014).
DOI: 10.1371/journal.ppat.1004245
Abstrakt: Parasitic nematodes are responsible for devastating illnesses that plague many of the world's poorest populations indigenous to the tropical areas of developing nations. Among these diseases is lymphatic filariasis, a major cause of permanent and long-term disability. Proteins essential to nematodes that do not have mammalian counterparts represent targets for therapeutic inhibitor discovery. One promising target is trehalose-6-phosphate phosphatase (T6PP) from Brugia malayi. In the model nematode Caenorhabditis elegans, T6PP is essential for survival due to the toxic effect(s) of the accumulation of trehalose 6-phosphate. T6PP has also been shown to be essential in Mycobacterium tuberculosis. We determined the X-ray crystal structure of T6PP from B. malayi. The protein structure revealed a stabilizing N-terminal MIT-like domain and a catalytic C-terminal C2B-type HAD phosphatase fold. Structure-guided mutagenesis, combined with kinetic analyses using a designed competitive inhibitor, trehalose 6-sulfate, identified five residues important for binding and catalysis. This structure-function analysis along with computational mapping provided the basis for the proposed model of the T6PP-trehalose 6-phosphate complex. The model indicates a substrate-binding mode wherein shape complementarity and van der Waals interactions drive recognition. The mode of binding is in sharp contrast to the homolog sucrose-6-phosphate phosphatase where extensive hydrogen-bond interactions are made to the substrate. Together these results suggest that high-affinity inhibitors will be bi-dentate, taking advantage of substrate-like binding to the phosphoryl-binding pocket while simultaneously utilizing non-native binding to the trehalose pocket. The conservation of the key residues that enforce the shape of the substrate pocket in T6PP enzymes suggest that development of broad-range anthelmintic and antibacterial therapeutics employing this platform may be possible.
Databáze: MEDLINE
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