Start Date
April 2024
Location
2nd floor - Library
Abstract
Green fluorescent protein (GFP) is a fluorescent protein due to the presence of a fluorophore produced from the cyclization of three residues (Ser65, Tyr66, and Gly67) located along an alpha helix surrounded by a beta-barrel. The goal of this study was to identify a non-fluorescent protein in the Protein Data Bank that had a similar three-residue motif that could be engineered to fluoresce as GFP does. Using a Structure Motif query in ASSAM, 1INL was identified as a strong candidate for this work. 1INL is a spermidine synthase from Thermotoga maritima. It is a tetramer, with each monomer consisting of many alpha helices. Examination of the structure of 1INL suggests that the lack of stabilizing residues around the motif prevents its cyclization into the fluorophore. Residues required for stabilizing the fluorophore will be inserted into 1INL by mutating the original residues to the corresponding stabilizing residues in GFP. The ability of the engineered protein to fluoresce will then be experimentally determined.
Creating light from darkness: engineering a non-fluorescent protein to fluoresce
2nd floor - Library
Green fluorescent protein (GFP) is a fluorescent protein due to the presence of a fluorophore produced from the cyclization of three residues (Ser65, Tyr66, and Gly67) located along an alpha helix surrounded by a beta-barrel. The goal of this study was to identify a non-fluorescent protein in the Protein Data Bank that had a similar three-residue motif that could be engineered to fluoresce as GFP does. Using a Structure Motif query in ASSAM, 1INL was identified as a strong candidate for this work. 1INL is a spermidine synthase from Thermotoga maritima. It is a tetramer, with each monomer consisting of many alpha helices. Examination of the structure of 1INL suggests that the lack of stabilizing residues around the motif prevents its cyclization into the fluorophore. Residues required for stabilizing the fluorophore will be inserted into 1INL by mutating the original residues to the corresponding stabilizing residues in GFP. The ability of the engineered protein to fluoresce will then be experimentally determined.
