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Sequence analysis of Plasmodium falciparum SERCA-type pfATPase6 in field isolates collected along Thai-Cambodian border Cover

Sequence analysis of Plasmodium falciparum SERCA-type pfATPase6 in field isolates collected along Thai-Cambodian border

Open Access
|Jan 2017

Figures & Tables

Figure 1

The crystal structure of the rabbit skeletal muscle SERCA1 showing the area around the thapsigargin-binding pocket composed of M3 helix (yellow), M5 helix (pink), and M7 helix (brown). The structure was modeled using the PyMOL Molecular Graphics System software (Schr dinger, New York, NY, USA) (available from www.pymol.org) to view the proposed catalytic site of the thapsigargin using the PDB entry 1IWO [25]. Artemisinin derivatives, which share some structural similarity with thapsigargin, were modeled and found to interact with the same regions of the Plasmodium falciparum ATPase6 protein [6, 8]. In addition, the majority of the coding sequence for these helices are conserved in the Plasmodium species, and the E255 in SERCA1 is a corresponding residue of the L263 in the pfATPase6 protein [6, 8].
The crystal structure of the rabbit skeletal muscle SERCA1 showing the area around the thapsigargin-binding pocket composed of M3 helix (yellow), M5 helix (pink), and M7 helix (brown). The structure was modeled using the PyMOL Molecular Graphics System software (Schr dinger, New York, NY, USA) (available from www.pymol.org) to view the proposed catalytic site of the thapsigargin using the PDB entry 1IWO [25]. Artemisinin derivatives, which share some structural similarity with thapsigargin, were modeled and found to interact with the same regions of the Plasmodium falciparum ATPase6 protein [6, 8]. In addition, the majority of the coding sequence for these helices are conserved in the Plasmodium species, and the E255 in SERCA1 is a corresponding residue of the L263 in the pfATPase6 protein [6, 8].

DNA sequences of the forward (F) and reverse (R) primers used in the PCR experiments

PrimerSequences
M3F5′-AGCATGCTGTTATAGAA-3′
M3R5′-TGAATTGGATCTGAGAAATGT-3′
M5F5′-TAATGGAACGGAGGTAGCTA-3′
M5R5′-ACGGGAGCTAAACTGTCAG-3′
M7F5′-TCCACCAGAACATGACGTAA-3′
M7R5′-AAAAACCAGTACACAAATATTGAGA-3′
769F5′-AAATATGGGAAAAAGACGATTAA-3′
769R5′-TACACGTATACCAGCCATATGG-3′

Sequence polymorphisms of pfATPase6 observed in diverse treatments and clinical outcome

RegimenGeographical RegionClinical OutcomeM3 Sequence Compared to 3D7 CloneSequence region around 769 residue Compared to 3D7 CloneM5 Sequence Compared to 3D7 CloneM7 Sequence Compared to 3D7 Clone
Artesunate monotherapyTasanh, CambodiaACPR (n = 70)Identical (n=70)Identical (n = 70)Identical (n = 46) Syn (n = 24)Identical (n = 70)
(n = 74)Failure (n = 4)
(D of treatment initiation, D0)Identical (n = 4)Identical (n = 4)Identical (n = 2) Syn (n = 2)Identical (n = 4)
Failure (n = 4)IdenticalIdentical (n = 4)Identical (n = 2)Identical
(D of failure Df)(n = 4)Syn (n = 2)(n = 4)
Quinine and tetracyclineTasanh, CambodiaACPR (n = 37)Identical (n = 37)Identical (n = 37)Identical (n = 26) Syn (n = 11)Identical (n = 37)
(n = 37)Failure (n = 0)
(D of treatment initiation, D0)N/AN/AN/AN/A
Failure (n = 0) (D of failure Df)N/AN/AN/AN/A
Artesunate and mefloquineTrat, Thailand (n = 53)ACPR (n = 44)Identical (n = 42) Unamp (n = 2)Identical (n = 42) Unamp (n = 2)Identical (n = 35) Syn (n = 7) Unamp (n = 2)Identical (n = 43) Unamp (n = 1)
Failure (n = 9)
(D of treatment initiation, D0)Identical (n = 9)Identical (n = 9)Identical (n = 9)Identical (n = 9)
Failure (n = 9) (D of failure Df)Identical (n = 9)Identical (n = 9)Identical (n = 5) Unamp (n = 4)Identical (n = 9)

Three regimens and clinical outcomes from two study sites along the Thai–Cambodian border

Study SitesAntimalarial TreatmentNumber of cases
ACPR

adequate clinical and parasitological response according to the WHO guidelines for the treatment of malaria assessed at 28 days for patients in Tasanh and 42 days for patients in Trat

Treatment failure

as defined by WHO in 2003

Tasanh, CambodiaArtesunate monotherapy 200 mg p.o. once daily × 7 d704
quinine 30 mg base/kg/day p.o. × 7 d; plus
tetracycline 25 mg/kg/day p.o. × 7d370
Trat, ThailandArtesunate 12 mg/kg p.o. once daily × 2 d; plus
mefloquine 25 mg/kg p.o. in 2 split doses; plus
primaquine 0.5mg/kg p.o. single dose449
Total15113
DOI: https://doi.org/10.5372/1905-7415.0903.403 | Journal eISSN: 1875-855X | Journal ISSN: 1905-7415
Language: English
Page range: 343 - 351
Published on: Jan 31, 2017
Published by: Chulalongkorn University
In partnership with: Paradigm Publishing Services
Publication frequency: 6 issues per year
Keywords:

© 2017 Wiriya Rutvisuttinunt, Kurt E. Schaecher, Harald Noedl, Chan Thap Lon, Stuart Tyner, Charlotte A. Lanteri, Krisada Jongsakul, Delia Bethell, Bryan L. Smith, David L. Saunders, Mark M. Fukuda, published by Chulalongkorn University
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.