2021 Student Research Conference:
34th Annual Student Research Conference

Determination of Re-Folding Effectiveness of DBF Through Lysozyme Activity Assays


Ryan P. Poland
Dr. Cassidy Dobson, Faculty Mentor

Cataracts are the result of improperly folded disulfide bonds, which form large aggregates in the eye called gamma crystallins. Disulfide Bond Forming enzyme (DBF) is known to correct improperly folded disulfide bonds, although it lacks cysteine residues in its amino acid sequence. The overall scope of this project is to elucidate a mechanism by which DBF re-folds disulfide bonds, and assess its effectiveness as a treatment for protein misfolding diseases such as cataracts. Using UV-Visible spectroscopy, lysozyme activity is monitored in the presence of DBF as a model. Denatured lysozyme is added to DBF, ATP/MgCl2, substrate, and buffer and allowed to react while absorbance data is collected. Variables such as [DBF], [substrate], and incubation time and temperature are changed independently in order to determine optimal reaction conditions. DBF test cases are compared to positive and negative controls using denatured or native lysozyme to assess the re-folding effectiveness of DBF.

 

Keywords: DBF, Lysozyme, Disulfide Bonds, Cataracts, Spectroscopy

Topic(s):Biochemistry and Molecular Biology
Chemistry
Biology

Presentation Type: Face-to-Face Oral Presentation

Session: 301-1
Location: SUB GEO
Time: 1:30

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