title: Probing function and structure of trehalose-6-phosphate phosphatases from pathogenic organisms suggests distinct molecular groupings.
abstract: The trehalose biosynthetic pathway is of great interest for the development of novel therapeutics because trehalose is an essential disaccharide in many pathogens but is neither required nor synthesized in mammalian hosts. As such, trehalose-6-phosphate phosphatase (TPP), a key enzyme in trehalose biosynthesis, is likely an attractive target for novel chemotherapeutics. Based on a survey of genomes from a panel of parasitic nematodes and bacterial organisms and by way of a structure-based amino acid sequence alignment, we derive the topological structure of monoenzyme TPPs and classify them into 3 groups. Comparison of the functional roles of amino acid residues located in the active site for TPPs belonging to different groups reveal nuanced variations. Because current literature on this enzyme family shows a tendency to infer functional roles for individual amino acid residues, we investigated the roles of the strictly conserved aspartate tetrad in TPPs of the nematodeBrugia malayiby using a conservative mutation approach. In contrast to aspartate-213, the residue inferred to carry out the nucleophilic attack on the substrate, we found that aspartate-215 and aspartate-428 ofBmTPP are involved in the chemistry steps of enzymatic hydrolysis of the substrate. Therefore, we suggest that homology-based inference of functionally important amino acids by sequence comparison for monoenzyme TPPs should only be carried out for each of the 3 groups.-Cross, M., Lepage, R., Rajan, S., Biberacher, S., Young, N. D., Kim, B.-N., Coster, M. J., Gasser, R. B., Kim, J.-S., Hofmann, A. Probing function and structure of trehalose-6-phosphate phosphatases from pathogenic organisms suggests distinct molecular groupings.
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Nathan, Queensland, Australia.
authors.forename: Megan
authors.initials: M
authors.lastname: Cross
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Nathan, Queensland, Australia.
authors.forename: Romain
authors.initials: R
authors.lastname: Lepage
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Nathan, Queensland, Australia.
authors.forename: Siji
authors.initials: S
authors.lastname: Rajan
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Nathan, Queensland, Australia.
authors.forename: Sonja
authors.initials: S
authors.lastname: Biberacher
authors.affiliation: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Neil D
authors.initials: ND
authors.lastname: Young
authors.affiliation: Department of Chemistry, Chonnam National University, Gwangju, South Korea; and.
authors.forename: Bo-Na
authors.initials: BN
authors.lastname: Kim
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Nathan, Queensland, Australia.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Coster
authors.affiliation: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Robin B
authors.initials: RB
authors.lastname: Gasser
authors.affiliation: Department of Chemistry, Chonnam National University, Gwangju, South Korea; and.
authors.forename: Jeong-Sun
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Eskitis Institute for Drug Discovery, Griffith University, Nathan, Queensland, Australia; a.hofmann@griffith.edu.au.|Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria, Australia.|Queensland Tropical Health Alliance, Smithfield, Queensland, Australia.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hofmann
mesh_terms: D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017178:Brugia malayi / Q000201:enzymology*; D020134:Catalytic Domain; D017124:Conserved Sequence; D015801:Helminth Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009161:Mycobacterium / Q000201:enzymology; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D015801:Helminth Proteins; D001224:Aspartic Acid; C023178:trehalose-phosphatase; D010744:Phosphoric Monoester Hydrolases
keywords: drug targets; enzyme inhibitors; molecular mechanism