Dr Denis Ptchelkine
Structural biology of DNA repair
Developing structural biology at WIMM is a new initiative that is driven by growing importance of understanding of functioning of proteins on molecular level for generation of new drugs and improvement of existing anti-cancer therapies.
Our work is focused on structural characterization of the protein complexes essential for the DNA repair and their components. We co-express the subunits of the complexes in our laboratory in the insect cells using MultiBac approach and we prepare high quality samples of the proteins on two HPLC AKTA systems which are part of a new mini-platform for protein purification that has recently been set up at the WIMM. We are gathering data on behavior of protein complexes in solution from a combination of biophysical methods such as SEC-MALS, AUC, ITC, SDS-PAGE, mass spectroscopy and SAXS. We are using cryo-electron microscopy and X-ray crystallography in order to achieve complete high resolution reconstruction of the key protein complexes involved in ICL repair. Our work benefits from established collaborations with other Departments of the University of Oxford (STRUBI, Dunn School of Pathology and Chemistry Department), Research Complex at Harwell and Diamond.
Bao L, Hannon C, Cruz-Migoni A, Ptchelkine D, Sun M, Derveni M, Bunjobpol W, Quevedo C, Miller A, Chambers J, Simmons A, Phillips S, Rabbitts T, Intracellular immunization against HIV infection with an intracellular antibody that mimics HIV integrase binding to cellular LEDGF protein. Scientific Reports 2017, 7(1): 16869
Hardwick J*, Ptchelkine D*, El-Sagheer A, Tear I, Singleton D, Phillips S, Lane A, Brown T. 5-Formylcytosine does not change the global structure of DNA. Nat Struct Biol 2017, 26(4), pp. 544-552
Ptchelkine D, Mochizuki T, Gillum A, Phillips SEV, Prangishvilli D, Huiskonen J, Unique architecture of thermophilic virus APBV1 and its genome packaging. Nat Commun 2017, 8(1): 1436
Abdullah UB, McGouran JF, Ptchelkine, D, El-Sagheer AH, Brown T and McHugh PJ RPA activates XPF-ERCC1 to initiate the processing of DNA interstrand crosslinks. EMBO J 2017, 36(14): 2047-2060.