Molecular modelling and kinase assay for CePNKP binding to DNA
Computational techniques of homology modelling, enzyme–substrate docking, and molecular dynamics were applied to elucidate the structure and substrate binding properties of the DNA repair enzyme polynucleotide kinase/phosphatase from the nematode C. elegans (CePNKP). PNKP is involved in the repair of DNA strand breaks, a form of DNA damage caused by reactive oxygen species, ionizing radiation and certain chemical mutagens. Mutations in human PNKP have been associated with the neurological disorders Microcephaly with Intractable Seizures (MCSZ) and Ataxia Oculomotor Apraxia 4 (AOA4). In addition, human PNKP has been identified as a potential drug target for the development of chemo- and radiosensitizing agents for cancer treatment. CePNKP is a useful model system for studying the human enzyme. The substrate preference for both human and C. elegans PNKP has been investigated, and found to be similar, with higher selectivity for recessed over blunt DNA ends. However, CePNKP exhibits a more exclusive preference for recessed DNA ends than human PNKP. To elucidate the reason for the unique substrate specificity of CePNKP, the structure of CePNKP in complex with its DNA substrate must be determined in atomistic detail by molecular modelling. The generated structural model is compared with our experimental results of kinase activity assays of wild-type CePNKP.
Presented on April 24, 2017 at Student Research Day held at MacEwan University in Edmonton, Alberta.
All Rights Reserved