The single cyclic nucleotide-specific phosphodiesterase of the intestinal parasite Giardia lamblia represents a potential drug target.

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  • Additional Information
    • Source:
      Publisher: Public Library of Science Country of Publication: United States NLM ID: 101291488 Publication Model: eCollection Cited Medium: Internet ISSN: 1935-2735 (Electronic) Linking ISSN: 19352727 NLM ISO Abbreviation: PLoS Negl Trop Dis Subsets: MEDLINE
    • Publication Information:
      Original Publication: San Francisco, CA : Public Library of Science
    • Subject Terms:
      Drug Discovery*; Antiprotozoal Agents/*pharmacology ; Giardia lamblia/*enzymology ; Giardia lamblia/*genetics ; Phosphodiesterase Inhibitors/*pharmacology ; Phosphoric Diester Hydrolases/*metabolism; Catalytic Domain ; Giardia lamblia/drug effects ; Giardia lamblia/growth & development ; Giardiasis/drug therapy ; Giardiasis/parasitology ; Humans ; Intestinal Diseases, Parasitic/drug therapy ; Intestinal Diseases, Parasitic/parasitology ; Phosphodiesterase Inhibitors/isolation & purification ; Phosphoric Diester Hydrolases/genetics ; Phosphoric Diester Hydrolases/isolation & purification ; Saccharomyces cerevisiae/genetics ; Sequence Alignment
    • Abstract:
      Background: Giardiasis is an intestinal infection correlated with poverty and poor drinking water quality, and treatment options are limited. According to the Center for Disease Control and Prevention, Giardia infections afflict nearly 33% of people in developing countries, and 2% of the adult population in the developed world. This study describes the single cyclic nucleotide-specific phosphodiesterase (PDE) of G. lamblia and assesses PDE inhibitors as a new generation of anti-giardial drugs.
      Methods: An extensive search of the Giardia genome database identified a single gene coding for a class I PDE, GlPDE. The predicted protein sequence was analyzed in-silico to characterize its domain structure and catalytic domain. Enzymatic activity of GlPDE was established by complementation of a PDE-deficient Saccharomyces cerevisiae strain, and enzyme kinetics were characterized in soluble yeast lysates. The potency of known PDE inhibitors was tested against the activity of recombinant GlPDE expressed in yeast and against proliferating Giardia trophozoites. Finally, the localization of epitope-tagged and ectopically expressed GlPDE in Giardia cells was investigated.
      Results: Giardia encodes a class I PDE. Catalytically important residues are fully conserved between GlPDE and human PDEs, but sequence differences between their catalytic domains suggest that designing Giardia-specific inhibitors is feasible. Recombinant GlPDE hydrolyzes cAMP with a Km of 408 μM, and cGMP is not accepted as a substrate. A number of drugs exhibit a high degree of correlation between their potency against the recombinant enzyme and their inhibition of trophozoite proliferation in culture. Epitope-tagged GlPDE localizes as dots in a pattern reminiscent of mitosomes and to the perinuclear region in Giardia.
      Conclusions: Our data strongly suggest that inhibition of G. lamblia PDE activity leads to a profound inhibition of parasite proliferation and that GlPDE is a promising target for developing novel anti-giardial drugs.
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    • Grant Information:
      R01 AI082577 United States AI NIAID NIH HHS
    • Accession Number:
      0 (Antiprotozoal Agents)
      0 (Phosphodiesterase Inhibitors)
      EC 3.1.4.- (Phosphoric Diester Hydrolases)
    • Publication Date:
      Date Created: 20170916 Date Completed: 20171023 Latest Revision: 20201214
    • Publication Date:
      20240829
    • Accession Number:
      PMC5617230
    • Accession Number:
      10.1371/journal.pntd.0005891
    • Accession Number:
      28915270