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Characterization of Plasmodium falciparum ATP-dependent DNA helicase RuvB3
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Metadata
Document Title
Characterization of Plasmodium falciparum ATP-dependent DNA helicase RuvB3
Author
Limudomporn P.,Moonsom S.,Leartsakulpanich U.,Suntornthiticharoen P.,Petmitr S.,Weinfeld M.,Chavalitshewinkoon-Petmitr P.
Name from Authors Collection
Affiliations
Department of Protozoology, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Bangkok, 10400, Thailand; National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Pahonyothin Rd, Pathumthani, 12120, Thailand; Department of Biomedical Sciences, Faculty of Science, Rangsit University, Lak Hok, Pathumthani, 12000, Thailand; Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400, Thailand; Department of Oncology, Cross Cancer Institute, University of Alberta, Edmonton, AB T6G 1Z2, Canada
Type
Article
Source Title
Malaria Journal
ISSN
14752875
Year
2016
Volume
15
Issue
1
Open Access
All Open Access, Gold, Green
Publisher
BioMed Central
DOI
10.1186/s12936-016-1573-2
Abstract
Background: Malaria is one of the most serious and widespread parasitic diseases affecting humans. Because of the spread of resistance in both parasites and the mosquito vectors to anti-malarial drugs and insecticides, controlling the spread of malaria is becoming difficult. Thus, identifying new drug targets is urgently needed. Helicases play key roles in a wide range of cellular activities involving DNA and RNA transactions, making them attractive anti-malarial drug targets. Methods: ATP-dependent DNA helicase gene (PfRuvB3) of Plasmodium falciparum strain K1, a chloroquine and pyrimethamine-resistant strain, was inserted into pQE-TriSystem His-Strep 2 vector, heterologously expressed and affinity purified. Identity of recombinant PfRuvB3 was confirmed by western blotting coupled with tandem mass spectrometry. Helicase and ATPase activities were characterized as well as co-factors required for optimal function. Results: Recombinant PfRuvB3 has molecular size of 59 kDa, showing both DNA helicase and ATPase activities. Its helicase activity is dependent on divalent cations (Cu2+, Mg2+, Ni+2 or Zn+2) and ATP or dATP but is inhibited by high NaCl concentration (>100 mM). PfPuvB3 is unable to act on blunt-ended duplex DNA, but manifests ATPase activity in the presence of either single- or double-stranded DNA. PfRuvB3.is inhibited by doxorubicin, daunorubicin and netropsin, known DNA helicase inhibitors. Conclusions: Purified recombinant PfRuvB3 contains both DNA helicase and ATPase activities. Differences in properties of RuvB between the malaria parasite obtained from the study and human host provide an avenue leading to the development of novel drugs targeting specifically the malaria form of RuvB family of DNA helicases. © 2016 The Author(s).
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
Funding Sponsor
Canadian Institutes of Health Research; Mahidol University; Thailand Research Fund
License
CC BY
Rights
Author
Publication Source
Scopus