2019 Student Research Conference:
32nd Annual Student Research Conference

Investigation of Gamma Crystallin Aggregation


Alexandra M. Marko
Dr. Cassidy Dobson, Faculty Mentor

Cataracts are caused by gamma crystallin aggregation and is the leading cause of blindness worldwide. Some gamma crystallin aggregates can form strong, intermolecular disulfide bonds. To study the possible reversal of aggregation, our lab seeks to understand the aggregation of the gamma crystallins over long periods of time. Recombinant Gamma S crystallin was expressed in E. coli cells and purified by IMAC. After purification, proteins were stored at 4°C to observe aggregation over time. Size Exclusion Column (SEC) was used to monitor protein aggregation and verified by SDS-PAGE. SEC chromatograms showed three possible Gamma S conformations: monomers, dimers, and aggregates. Older proteins showed a possible equilibrium between species while newer samples showed distinct species. Studying the aggregation of gamma crystallin proteins will assist our lab in understanding the behavior of an enzyme, Disulfide Bond Forming enzyme (DBF), and the possible reversal of gamma crystallin aggregation.

Keywords: gamma crystallins, protein aggregation, SEC, SDS-PAGE

Topic(s):Biochemistry and Molecular Biology
Biology
Chemistry

Presentation Type: Oral Paper

Session: 211-2
Location: MC 212
Time: 10:30

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