A novel protein footprinting platform: mass spectrometry of laser-initiated carbene reactions

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dc.contributor.advisorSchriemer, David C.
dc.contributor.authorJumper, Chanelle C.
dc.date.accessioned2017-12-18T22:16:39Z
dc.date.available2017-12-18T22:16:39Z
dc.date.issued2011
dc.descriptionBibliography: p. 114-123en
dc.descriptionA few pages are in colour.en
dc.descriptionIncludes copies of copyright permission. Original copies with original Partial Copyright Licence.en
dc.description.abstractThis work reports a protein labeling method using non-selective carbene reactions of sufficiently high efficiency to permit detection by mass spectrometric methods at the protein, peptide and residue level. The approach uses a diazirine-modified amino acid (L-2-amino-4,4' -azipentanoic acid, "photoleucine") as a label source, which is converted to a highly reactive carbene by pulsed laser photolysis at 355 nm. Labeling of model proteins and peptides was achieved with sensitivity to changes in protein topography brought about by conformational change and ligand binding. Labeling yield is independent of protein concentration over approximately two orders of magnitude, but is weakly dependent on the presence of other chromophores in a photon-limited apparatus. The current configuration required 2 minutes of irradiation for full reagent conversion, however it is shown that comparable yields can be achieved with a single high-energy laser pulse (>100 mJ/pulse, <10 nsec), offering a labeling method with high temporal resolution.
dc.format.extentxiv, 124 leaves : ill. ; 30 cm.en
dc.identifier.citationJumper, C. C. (2011). A novel protein footprinting platform: mass spectrometry of laser-initiated carbene reactions (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/3998en_US
dc.identifier.doihttp://dx.doi.org/10.11575/PRISM/3998
dc.identifier.urihttp://hdl.handle.net/1880/104999
dc.language.isoeng
dc.publisher.institutionUniversity of Calgaryen
dc.publisher.placeCalgaryen
dc.rightsUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.
dc.titleA novel protein footprinting platform: mass spectrometry of laser-initiated carbene reactions
dc.typemaster thesis
thesis.degree.disciplineChemistry
thesis.degree.grantorUniversity of Calgary
thesis.degree.nameMaster of Science (MSc)
ucalgary.item.requestcopytrue
ucalgary.thesis.accessionTheses Collection 58.002:Box 2019 627942869
ucalgary.thesis.notesUARCen
ucalgary.thesis.uarcreleaseyen
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