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Use of a Paramagnetic Spin Label for Determination of Long-Range Distance Constraints in Solid-State NMR

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dc.contributor.advisor Ladizhansky, Vladimir
dc.contributor.advisor Brown, Leonid O'Halloran, Matthew 2014-08-12T18:01:16Z 2014-08-12T18:01:16Z 2014-07 2014-07-28 2014-08-12
dc.description.abstract Solid-State Nuclear Magnetic Resonance (SSNMR), with paramagnetic relaxation enhancement (PRE), is used to investigate the protein Proteorhodopsin (PR). PRE allows for the acquisition of long-range distance constraints not accessible through standard NMR experiments. Four mutant versions of PR, created to have only one cysteine residue in each, were investigated. The S215C mutant did not purify well and appeared to retain a large amount of detergent. The S55C variant was found to differ significantly from the wild-type structure. The C107 mutant experienced delocalized PRE effects originally thought to be due to proton spin diffusion, when this interaction was removed to the first order using Lee-Goldburg cross-polarization (CP), the delocalized effect was still observed. The C175 mutant produced long-range distance restraints that can be used to restrain three of the helices in the bundle on the cytoplasmic side. Future studies are required to better understand the structure of this membrane protein. en_US
dc.description.sponsorship NSERC PGS M, Ontario Graduate Scholarship (OGS) Program en_US
dc.language.iso en en_US
dc.publisher University of Guelph en_US
dc.rights Attribution-NonCommercial-NoDerivs 2.5 Canada *
dc.rights.uri *
dc.subject Solid State Nuclear Magnetic Resonance (SSNMR) en_US
dc.subject Paramagnetic Relaxation Enhancement (PRE) en_US
dc.subject Membrane Proteins en_US
dc.subject Proteorhodopsin (PR) en_US
dc.subject Site Specific Labelling en_US
dc.title Use of a Paramagnetic Spin Label for Determination of Long-Range Distance Constraints in Solid-State NMR en_US
dc.type Thesis en_US Biophysics en_US Master of Science en_US Department of Physics en_US University of Guelph en_US

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Attribution-NonCommercial-NoDerivs 2.5 Canada Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 2.5 Canada