Prevention of Gamma Crystallin Aggregation with DBF

Alexandra M. Marko
Dr. Cassidy Dobson, Faculty Mentor

Cataracts are the leading cause of blindness worldwide, and some forms are caused by strong, intermolecular disulfide bonds between gamma crystallin protein aggregates. Our lab seeks to understand if Disulfide Bond Forming enzyme (DBF) can act as a molecular chaperone to prevent the aggregation of gamma crystallins. Recombinant Gamma S crystallin was expressed in E. coli cells and purified by Nickel IMAC. Gamma crystallins were incubated for two days in the presence and absence of DBF, ATP, and MgCl2. Size Exclusion Column (SEC) was used to monitor protein aggregation in the presence and absence of DBF. SEC chromatograms showed no difference between Gamma S in the presence and absence of DBF. Gamma S showed no characteristic alterations in the presence of ATP and MgCl2. Future experiments will test more concentrated gamma crystallins in order to quantitatively and qualitatively assess the impact of DBF on gamma crystallin aggregation.

Keywords: gamma crystallins, Disulfide Bond Forming enzyme, protein aggregation, SEC

Topic(s):Biochemistry and Molecular Biology

Presentation Type: Oral Presentation

Session: TBA
Location: TBA
Time: TBA