Molecular characterization and localization of Plasmodium falciparum choline kinase

Biochimica Et Biophysica Acta
Vinay ChoubeyUday Bandyopadhyay

Abstract

Generation of phosphocholine by choline kinase is important for phosphatidylcholine biosynthesis via Kennedy pathway and phosphatidylcholine biosynthesis is essential for intraerythrocytic growth of malaria parasite. A putative gene (Gene ID PF14_0020) in chromosome 14, having highest sequence homology with choline kinase, has been identified by BLAST searches from P. falciparum genome sequence database. This gene has been PCR amplified, cloned, over-expressed and characterized. Choline kinase activity of the recombinant protein (PfCK) was validated as it catalyzed the formation of phosphocholine from choline in presence of ATP. The K(m) values for choline and ATP are found to be 145+/-20 microM and 2.5+/-0.3 mM, respectively. PfCK can phosphorylate choline efficiently but not ethanolamine. Southern blotting indicates that PfCK is a single copy gene and it is a cytosolic protein as evidenced by Western immunoblotting and confocal microscopy. A model structure of PfCK was constructed based on the crystal structure of choline kinase of C. elegans to search the structural homology. Consistent with the homology modeling predictions, CD analysis indicates that the alpha and beta content of PfCK are 33% and 14%, respectively. Since c...Continue Reading

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Citations

Dec 6, 2006·Antimicrobial Agents and Chemotherapy·Vinay ChoubeyUday Bandyopadhyay
Sep 4, 2010·Molecular and Biochemical Parasitology·Gregory J CrowtherKelli L Kuhen
Jul 6, 2010·International Journal for Parasitology·Sandrine DéchampsHenri J Vial
Dec 19, 2009·Analytical Biochemistry·Gregory J CrowtherWesley C Van Voorhis
May 6, 2008·Molecular and Biochemical Parasitology·Shweta JoshiJitendra Kumar Saxena
Dec 12, 2007·Free Radical Biology & Medicine·Sanjay KumarUday Bandyopadhyay
Dec 22, 2006·Protein Expression and Purification·Mithu GuhaUday Bandyopadhyay
May 4, 2016·Antimicrobial Agents and Chemotherapy·Souvik SarkarUday Bandyopadhyay
Sep 1, 2009·Expert Review of Clinical Pharmacology·Athar AlamUday Bandyopadhyay
Jul 23, 2020·BioMed Research International·Moad KhalifaWei Cun See Too
Feb 2, 2021·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Pooja Rani MinaMahendra P Darokar
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Mar 30, 2021·Frontiers in Pharmacology·Saurabh KumarMahendra P Darokar
Jun 3, 2021·Pharmaceutics·Juan Carlos LacalJoaquín M Campos

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