Ng, KennethArellano Saab, Amir Alam2016-08-192016-08-1920162016http://hdl.handle.net/11023/3192A variant of streptavidin, SAVSBPM32, was previously designed to incorporate mutations that weaken binding to biotin and introduce a cysteine at position 86. Streptavidin-binding peptides have also been re-designed to test the binding capabilities of SAVSBPM32. The primary goal of this project was to crystallize SAVSBPM32 and evaluate its interactions with biotin, a cysteine containing SBP-Tag and a cysteine containing a biotinylation tag (CPFB-2). The crystal structure of SAVSBPM32 in complex with biotin revealed for the first time that the loop bridging the third and fourth b-strands in SAVSBPM32 can adopt a closed or an open conformation. The crystal structure of SAVSBPM32 in complex with CPFB-2 reveals the novel structure of a biotinylated peptide bound to streptavidin while also forming a disulphide bond. These findings confirm key design principles and suggest approaches for further development.engUniversity 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.BiologyBiology--MolecularBiochemistryStreptavidinCrystallographyBiochemistryMutationStructuremaster thesis10.11575/PRISM/25498