34th Annual Student Research Conference
Gamma Crystallin Aggregation in the Presence of Disulfide Bond Forming (DBF) Enzyme
Alexandra M. Marko
Dr. Cassidy Dobson, Faculty Mentor
Cataracts are the leading cause of blindness worldwide, and a portion of cataracts are caused by strong, intermolecular disulfide bonds between gamma crystallin aggregates. Our lab seeks to study if Disulfide Bond Forming (DBF) Enzyme can be used as a molecular chaperone to prevent the formation of disulfide-mediated aggregates. In this study, recombinant Gamma S crystallin was expressed in E. coli and purified by IMAC. Purified gamma S was incubated at 37°C in the presence and absence of DBF. Aggregation patterns were qualitatively observed over time using SDS-PAGE. We predict there will be increased amounts of monomers and reduced amounts of dimers, trimers, and aggregates in the Gamma S/DBF experimental tube compared to the Gamma-S only control tube. Future experiments will study if DBF can be used to reverse Gamma S aggregates and use Size Exclusion Chromatography to quantitatively assess DBF’s impact on gamma crystallin aggregation.
Keywords: gamma crystallins, disulfide bond forming enzyme, DBF, protein aggregation, SDS-PAGE
Topic(s):Biology
Biochemistry and Molecular Biology
Chemistry
Presentation Type: Asynchronous Virtual Oral Presentation
Session: 3-2
Location: https://flipgrid.com/f86d186b
Time: 0:00