Experimental parasitology, 2013-07, Vol.134 (3), p.362-367
2013
Volltextzugriff (PDF)
Details
- Autor(en) / Beteiligte
- Titel
- Molecular and biochemical characterisation of arginine kinases in Haemonchus contortus and Teladorsagia circumcincta
- Ist Teil von
- Experimental parasitology, 2013-07, Vol.134 (3), p.362-367
- Ort / Verlag
- United States: Elsevier Inc
- Erscheinungsjahr
- 2013
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- •T. circumcincta and H. contortus arginine kinase genes (1080bp) encoded 360 amino acid proteins.•TcAK and HcAK proteins had 99% similarity with each other and 94% similarity to C. elegans AK.•Kinetic properties of recombinant TcAK and HcAK were very similar.•Arginine analogues strongly inhibited enzyme activity. Full length cDNA encoding arginine kinases (AK) were cloned from Teladorsagia circumcincta (TcAK) and Haemonchus contortus (HcAK). The TcAK and HcAK cDNA (1080bp) encoded 360 amino acid proteins. The predicted amino acid sequence showed 99% similarity with each other and 94% with a Caenorhabditis elegans AK. Soluble N-terminal His-tagged AK proteins were expressed in Escherichia coli strain BL21, purified and characterised. All binding sites were completely conserved in both proteins. The recombinant TcAK and HcAK had very similar kinetic properties: Km arginine was 0.35mM, Km ATP was 0.8–0.9mM and the pH optima were pH 7.5. Arginine analogues strongly inhibited recombinant enzyme activities (up to 80%), whilst other amino acids decreased activities by a maximum of 20%. TcAK and HcAK are potential vaccine candidates because of the strong antigenicity of invertebrate phosphagens and kinases and presence in metabolically active parts of the worm.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0014-4894eISSN: 1090-2449DOI: 10.1016/j.exppara.2013.03.011Titel-ID: cdi_proquest_miscellaneous_1365052869
- Format
- –
- Schlagworte
- adenosine triphosphate, Amino Acid Sequence, amino acid sequences, Animals, arginine, Arginine - metabolism, Arginine kinase, Arginine Kinase - antagonists & inhibitors, Arginine Kinase - chemistry, Arginine Kinase - genetics, Binding Sites, Caenorhabditis elegans, Cloning, Molecular, complementary DNA, DNA, Complementary - genetics, DNA, Helminth - genetics, Electrophoresis, Polyacrylamide Gel, enzyme activity, Escherichia coli, Gene sequence, Haemonchus - enzymology, Haemonchus - genetics, Haemonchus contortus, Helminth Proteins - chemistry, Helminth Proteins - genetics, Hydrogen-Ion Concentration, invertebrates, Kinetic properties, Kinetics, Molecular Sequence Data, parasitology, phosphotransferases (kinases), protein synthesis, proteins, Recombinant Proteins - chemistry, Recombinant Proteins - genetics, Sequence Alignment, Teladorsagia circumcincta, Trichostrongyloidea - enzymology, Trichostrongyloidea - genetics, vaccines