Monitoring Lysozyme Activity to Assess Re-Folding Effectiveness of DBF

Ryan P. Poland
Dr. Cassidy Dobson, Faculty Mentor

Cataracts are the leading cause of blindness worldwide, and are the result of incorrectly paired disulfide bonds, ultimately forming gamma crystallin aggregates. The overall goal of this research lab is to understand the effectiveness of Disulfide Bond Forming enzyme (DBF) as a molecular chaperone that refolds disulfide bonds. One way to model the effectiveness of DBF is to monitor the activity of denatured lysozyme in the presence of DBF. Using UV-Visible spectroscopy, the activity of lysozyme is observed over time, and changing parameters such incubation temperature, time, [ATP], and [DBF] gives insight into the conditions required for DBF to refold disulfide bonds. The goal of this assay is to determine the effect of incubation temperature on how well DBF can refold lysozyme’s intramolecular disulfide bonds. In future experiments, more incubation temperatures will be tested to determine the optimal temperature required for DBF refolding.

 

Keywords: DBF, Lysozyme, Disulfide Bonds

Topic(s):Biochemistry and Molecular Biology
Chemistry

Presentation Type: Oral Presentation

Session: TBA
Location: TBA
Time: TBA