Molecular cloning, expression, characterization and mutation of Plasmodium falciparum guanylate kinase
| Autor: | Tarek Yosef, Mahmoud Kandeel, Yoshihito Ueno, Masayuki Nakanishi, Takayuki Ando, Kamal El-Shazly, Yukio Kitade |
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| Rok vydání: | 2007 |
| Předmět: |
Guanylate kinase
Mutant DNA Mutational Analysis Molecular Sequence Data Plasmodium falciparum Guanosine Monophosphate Mutation Missense Gene Expression Sequence alignment Molecular cloning Adenosine Triphosphate PlasmoDB Animals Amino Acid Sequence Kinase activity Cloning Molecular Enzyme Inhibitors Site-directed mutagenesis Molecular Biology biology Deoxyguanine Nucleotides biology.organism_classification Molecular biology Kinetics Biochemistry Amino Acid Substitution Parasitology Guanylate Kinases Sequence Alignment |
| Zdroj: | Molecular and biochemical parasitology. 159(2) |
| ISSN: | 0166-6851 |
| Popis: | The present work describes cloning, expression, purification, characterization, and mutation of Plasmodium falciparum guanylate kinase (PlasmoDB ID PFI1420w). Amino-acid sequence alignment revealed important differences especially in K42-V51, Y73-A77, and F100-L110, which include residues important for kinase activity, and at helix 3, which is important for domain movements. The catalytic efficiency for dGMP was 22-fold lower than that for GMP, whose value is the lowest among known guanylate kinases. dGMP was found to a competitive inhibitor for GMP with Ki = 0.148 mM and a mixed-type inhibitor with regard to ATP with measured Ki = 0.4 mM. The specificity constant (Kcat/Km) of the four examined mutants varied for natural substrate GMP/dGMP, indicating the involvement of different mechanisms in substrate recognition and subsequent loop-domain movement. These results show that P. falciparum guanylate kinase is structurally and biochemically distinct from other guanylate kinases and could be a possible target in drug development. |
| Databáze: | OpenAIRE |
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