文章基本信息

标题：Bioinformatics, Enzymologic Properties, and Comprehensive Tracking of Plasmodium falciparum Nucleoside Diphosphate Kinase
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作者：Mahmoud Kandeel ; Tatsuya Miyamoto ; Yukio Kitade 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2009
卷号：32
期号：8
页码：1321-1327
DOI：10.1248/bpb.32.1321
出版社：The Pharmaceutical Society of Japan
摘要：The gene encoding for nucleoside diphosphate kinase from Plasmodium falciparum was obtained by polymerase chain reaction (PCR) and expressed in Escherichia coli . Tracking kinases is strenuous work due to many functional and technical deficits. Tracking of Plasmodium falciparum nucleoside diphosphate kinase (PfNDK) was carried out by conventional enzyme assays combined by isothermal titration calorimetry (ITC). ITC proved an efficient tracking method with rapid, accurate, and confident target confirmation. In addition, it provides substrate affinity and full thermodynamic profile in one experiment. Magnesium ions were found to be essential for nucleoside diphosphate (NDP) kinase activity; however, the absence of Mg²⁺ did not completely interfere with the binding of nucleotides. The substrate recognition was found to depend on enthalpic forces with little entropic contributions. However, in the absence of magnesium ions the nucleotides actively bind to the enzyme driven by hydrophobic forces. The enzyme showed specific activity that was within the average of known enzymes; however, it was at least two-fold higher than that of the human enzyme.
关键词：nucleoside diphosphate kinase;Plasmodium falciparum;isothermal titration calorimetry