Structure and dynamics of the Ovarian Tumour Domain Protease from the Crimean-Congo Hemorrhagic Fever Virus by Nuclear Magnetic Resonance spectroscopy
| dc.contributor.author | Saran, Sagar | |
| dc.contributor.examiningcommittee | Dibrov, Pavel (Microbiology) McKenna, Sean (Chemistry) | en_US |
| dc.contributor.supervisor | O'Nei, Joe (Chemistry) | en_US |
| dc.date.accessioned | 2016-09-15T20:25:03Z | |
| dc.date.available | 2016-09-15T20:25:03Z | |
| dc.date.issued | 2016 | |
| dc.degree.discipline | Chemistry | en_US |
| dc.degree.level | Master of Science (M.Sc.) | en_US |
| dc.description.abstract | Crimean-Congo Hemorrhagic Fever Virus (CCHFV) is endemic to more than 30 countries and shows fatality rates among humans ranging from 30-50%. The CCHFV 169-residue L-segment deubiquitinase (DUB) is a member of the superfamily of Ovarian Tumour (OTU) ubiquitin thiolesterases that interfere with innate immune responses and hence is an attractive antiviral target. I report here the application of Nuclear Magnetic Resonance spectroscopy to probe the role of CCHFV OTU enzyme dynamics in the catalytic mechanism of the enzyme. 13C/15N triple-resonance experiments and an amino acid “unlabelling” scheme were used for backbone resonances assignments. NMR chemical shift analysis, NMR spin-relaxation experiments at two magnetic fields, Lipari-Szabo Model-free formalism, reduced spectral density mapping and Carr-Purcell-Meiboom-Gill Relaxation Dispersion experiments were done to obtain structure and dynamics data. The dynamics data suggested an unfolded C-terminus and a well-packed protein core. Relaxation dispersion measurements show that a significant number of residues undergo conformational exchange on the millisecond timescale. Some of these are near the active site and neighbouring segments and may represent a rate-limiting event along the proteolytic kinetic pathway. They may also play a role in determining the enzyme’s broad substrate specificity. | en_US |
| dc.description.note | October 2016 | en_US |
| dc.identifier.uri | http://hdl.handle.net/1993/31783 | |
| dc.language.iso | eng | en_US |
| dc.rights | open access | en_US |
| dc.subject | Crimean-Congo Hemorrhagic Fever, Enzyme, NMR spectroscopy, Protease, Protein, Protein dynamics, Protein structure, Virus, | en_US |
| dc.title | Structure and dynamics of the Ovarian Tumour Domain Protease from the Crimean-Congo Hemorrhagic Fever Virus by Nuclear Magnetic Resonance spectroscopy | en_US |
| dc.type | master thesis | en_US |