Expression and Purification of Disulfide Bond Forming Enzyme

Mary C. Butler
Dr. Cassidy Dobson, Faculty Mentor

Aggregation of proteins is responsible for many neurodegenerative disorders such as Alzheimer’s and Amyotrophic Lateral Sclerosis (ALS).  Incorrect disulfide bond interactions can lead to protein aggregates that ultimately lead to protein misfolding diseases, such as cataracts. Disulfide bond forming enzyme (DBF) is a small molecular chaperone, able to rearrange incorrectly made disulfide bonds. This is comparable to Protein Disulfide Isomerase (PDI) which has a molecular weight of 57 kDa and has a similar function. DBF lacks internal cysteines, normally responsible for the activity of PDI. In order to study this unique mechanism, purified protein must first be obtained. DBF is expressed in E. coli and purified by nickel chromatography and analyzed using SDS-PAGE. Future work will be done to determine DBF’s mechanism of disulfide bond rearrangement by measuring its activity.  

Keywords: DBF, SDS-PAGE, protein purification

Topic(s):Biochemistry and Molecular Biology
Chemistry
Biology

Presentation Type: Oral Presentation

Session: TBA
Location: TBA
Time: TBA