Interaction of tubulin and protein kinase CK2 in Trypanosoma equiperdum
| Autor: | Beatriz E. Boscán, Graciela L. Uzcanga, Frank Aponte, José Bubis, Maritza Calabokis, Rocío Camargo |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Trypanosoma Emodin GTP' medicine.drug_class Immunoprecipitation 030231 tropical medicine Static Electricity Protozoan Proteins macromolecular substances Monoclonal antibody General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 0302 clinical medicine Tubulin medicine Phosphorylation Casein Kinase II Polyacrylamide gel electrophoresis Protein Kinase Inhibitors biology Chemistry Kinase Antibodies Monoclonal biology.organism_classification Molecular Weight Protein Transport 030104 developmental biology Biochemistry Gene Expression Regulation biology.protein Trypanosoma equiperdum Guanosine Triphosphate Protein Binding Signal Transduction |
| Zdroj: | Zeitschrift fur Naturforschung. C, Journal of biosciences. 72(11-12) |
| ISSN: | 0939-5075 |
| Popis: | A polypeptide band with an apparent molecular weight of 55,000 was phosphorylated in vitro in whole-cell lysates of Trypanosoma equiperdum. This band corresponds to tubulin as demonstrated by immunoprecipitation of the phosphorylated polypeptide from T. equiperdum extracts when anti-α and anti-β tubulin monoclonal antibodies were employed. A parasite protein kinase CK2 was in charge of modifying tubulin given that common mammalian CK2 inhibitors such as emodin and GTP, hindered the phosphorylation of tubulin and exogenously added casein. Interestingly, a divalent cation-dependent translocation of the T. equiperdum tubulin and the CK2 responsible for its phosphorylation was noticed, suggesting a direct interaction between these two proteins. Additionally, this fraction of tubulin and its kinase coeluted using separations based on parameters as different as charge (DEAE-Sepharose anion-exchange chromatography) and size (Sephacryl S-300 gel filtration chromatography). Analyses by non-denaturing polyacrylamide gel electrophoresis and immunoblot of the purified and radioactively labeled fraction containing both tubulin and the CK2 enzyme, established the phosphorylation of a single band that was recognized by anti-CK2 α-subunit and anti-tubulin antibodies. All these findings revealed a physical association between a pool of tubulin and a CK2 in T. equiperdum. |
| Databáze: | OpenAIRE |
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