Open AccessExpression, purification, crystallization and preliminary X‐ray analysis of Plasmodium falciparum GTP:AMP phosphotransferase
Author(s) -
Law Alan W. L.,
Lescar Julien,
Hao Quan,
Kotaka Masayo
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112015862
Subject(s) - phosphotransferase , gtp' , adenylate kinase , biochemistry , enzyme , crystallization , substrate (aquarium) , plasmodium falciparum , escherichia coli , nucleotide , kinase , guanosine triphosphate , chemistry , biology , gene , organic chemistry , ecology , malaria , immunology
Adenylate kinases (AKs) are phosphotransferase enzymes that catalyze the interconversion of adenine nucleotides, thereby playing an important role in energy metabolism. In Plasmodium falciparum , three AK isoforms, namely Pf AK1, Pf AK2 and GTP:AMP phosphotransferase ( Pf GAK), have been identified. While Pf AK1 and Pf AK2 catalyse the conversion of ATP and AMP to two molecules of ADP, Pf GAK exhibits a substrate preference for GTP and AMP and does not accept ATP as a substrate. Pf GAK was cloned and expressed in Escherichia coli and purified using two‐step chromatography. Brown hexagonal crystals of Pf GAK were obtained and a preliminary diffraction analysis was performed. X‐ray diffraction data for a single Pf GAK crystal were processed to 2.9 Å resolution in space group P 3 1 21 or P 3 2 21, with unit‐cell parameters a = b = 123.49, c = 180.82 Å, α = β = 90, γ = 120°.