Detection of the in vitro modulation of Plasmodium falciparum Arf1 by Sec7 and ArfGAP domains using a colorimetric plate-based assay.

Scientific Reports
Tarryn SwartHeinrich C Hoppe

Abstract

The regulation of human Arf1 GTPase activity by ArfGEFs that stimulate GDP/GTP exchange and ArfGAPs that mediate GTP hydrolysis has attracted attention for the discovery of Arf1 inhibitors as potential anti-cancer agents. The malaria parasite Plasmodium falciparum encodes a Sec7 domain-containing protein - presumably an ArfGEF - and two putative ArfGAPs, as well as an Arf1 homologue (PfArf1) that is essential for blood-stage parasite viability. However, ArfGEF and ArfGAP-mediated activation/deactivation of PfArf1 has not been demonstrated. In this study, we established an in vitro colorimetric microtiter plate-based assay to detect the activation status of truncated human and P. falciparum Arf1 and used it to demonstrate the activation of both proteins by the Sec7 domain of ARNO, their deactivation by the GAP domain of human ArfGAP1 and the inhibition of the respective reactions by the compounds SecinH3 and QS11. In addition, we found that the GAP domains of both P. falciparum ArfGAPs have activities equivalent to that of human ArfGAP1, but are insensitive to QS11. Library screening identified a novel inhibitor which selectively inhibits one of the P. falciparum GAP domains (IC50 4.7 µM), suggesting that the assay format is sui...Continue Reading

References

Jul 1, 1992·Molecular and Biochemical Parasitology·J L Crary, K Haldar
Nov 1, 1994·Experimental Parasitology·S A Ogun, A A Holder
Feb 1, 1993·The American Journal of Tropical Medicine and Hygiene·M T Makler, D J Hinrichs
Feb 1, 1997·Molecular and Biochemical Parasitology·R M TruongD E Goldberg
Aug 1, 1997·Molecular and Biochemical Parasitology·F J LeeC Syin
May 3, 2003·Nature Reviews. Molecular Cell Biology·Juan S Bonifacino, Jennifer Lippincott-Schwartz
Jan 27, 2004·Molecular Biology of the Cell·Randal CoxNava Segev
Apr 25, 2006·Nature Reviews. Molecular Cell Biology·Crislyn D'Souza-Schorey, Philippe Chavrier
Dec 15, 2006·Nature·Markus HafnerMichael Famulok
Apr 27, 2007·Proceedings of the National Academy of Sciences of the United States of America·Qisheng ZhangSheng Ding
Jun 15, 2007·Proceedings of the National Academy of Sciences of the United States of America·Julien ViaudAlain Chavanieu
Jan 28, 2009·International Journal for Parasitology·Werner LeberDavid A Baker
Feb 3, 2009·Nature Chemical Biology·José B SáenzDavid B Haslam
Jul 28, 2009·FEBS Letters·R BeckD Cassel
Feb 10, 2010·Molecular Microbiology·Thomas D OttoManuel Llinás
Apr 17, 2010·FEBS Letters·Anne SpangPaul A Randazzo
Sep 21, 2010·Current Protocols in Cell Biology·Lee Ann Cohen, Julie G Donaldson
May 19, 2011·Nature Reviews. Molecular Cell Biology·Julie G Donaldson, Catherine L Jackson
Oct 10, 2012·Molecular and Biochemical Parasitology·Marcel DeponteJude M Przyborski
Dec 27, 2012·Bioorganic & Medicinal Chemistry·Monika RajParamjit S Arora
Jun 29, 2013·Journal of Medicinal Chemistry·Kai SeehaferChristian D Klein
Aug 26, 2014·Journal of Cell Science·Catherine L Jackson, Samuel Bouvet
Sep 23, 2014·Chemistry & Biology·Michelle R ArkinJames A Wells
Apr 25, 2015·PloS One·Thuvaraka ThavayogarajahAnthony A Holder
Jul 15, 2015·Bioorganic & Medicinal Chemistry Letters·Manish K SinghQisheng Zhang
Sep 3, 2016·Small GTPases·Cristina CasalouDuarte C Barral
Mar 29, 2018·Parasites & Vectors·Holly MatthewsCatherine J Merrick
May 15, 2018·ChemMedChem·Mayibongwe J LungaClinton G L Veale
Mar 20, 2019·Cells·Néstor Prieto-DominguezYong Teng
May 16, 2019·Molecular Biology of the Cell·Elizabeth SztulRichard A Kahn

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Methods Mentioned

BETA
GTPases
nucleotide
GTPase
nucleotide exchange
PCR
pull-down
myristoylation
FRET
transgenic
protein assay

Software Mentioned

GraphPad Prism

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