Discipline: Chemistry and Chemical Sciences
Subcategory: Biochemistry (not Cell and Molecular Biology and Genetics)
Chely Tejeda - California State University, Los Angeles
Co-Author(s): Rebecca Vargas, California State University, Los Angeles, Los Angeles, CA.
Protein aggregation is biological phenomenon that occurs due to the misfolding or unfolding of proteins. This phenomenon is strongly associated with neurodegenerative diseases such as Alzheimer’s, Parkinson’s, and Huntington’s disease (Ross & Poirier, 2004). Protein aggregation is also a major issue in drug development. Recent results have sparked a concern about the presence of therapeutic protein aggregation in pharmaceuticals causing patients to become drug-tolerant (Roberts, 2014). Overall, the concept of anti-aggregation is not entirely understood and becoming more knowledgeable about the prevention of this occurrence can be helpful in creating effective biotherapies as well as gain more insight on disease caused by protein denaturation and aggregation. My study will investigate if Dendroides canadensis antifreeze protein 1 (DAFP-1) has protective characteristics against enzymes that are susceptible to denaturation and aggregation. Specifically, the enzymes that will be studied are lactate dehydrogenase (LDH) and alcohol dehydrogenase (ADH), which are both commonly present in many organisms and important for metabolism. DAFP-1 consists of eight disulfide bonds and is stable at high temperatures. On the other hand, LDH and ADH readily denature and loose enzymatic activity when heat-treated. Results show that DAFP-1 can significantly protect the enzymatic activity of LDH and ADH at 40 °C and 46 °C for about 21 minutes. To continue this study, the next steps are to quantify the degree of protection and anti-aggregation that DAFP-1 has on LDH and ADH when heat-treated. To do so, an aggregation assay and protein activity gel will be conducted. These methods will allow us to calculate the amount of LDH and ADH aggregation in the absence and presence of DAFP-1 due to the increasing temperature treatments, as well as determine if LDH and ADH retain their enzymatic activity after heat-treatment. We hypothesize that when exposed to heat, both LDH and ADH will have more enzymatic activity and less aggregation in the presence of DAFP-1 than in the absence of DAFP-1. Based on the data collected from these experiments, the protective and anti-aggregation characteristics of DAFP-1 will be further determined.Not Submitted
Funder Acknowledgement(s): National Science Foundation Grant #: HRD-1700556
Faculty Advisor: Dr. Xin Wen, firstname.lastname@example.org
Role: As a new Master's student, I have successfully grown and purified DAFP-1. My plans are to repeat the enzyme activity assays to determine the effects of DAFP-1 on LDH and ADH when heated and further support Rebecca's results. I plan to collect data that shows this effect over a consecutive range of temperatures. Also, I plan to use Thioflavin T dye to quantify the amount of LDH and ADH aggregation induced by the heat treatments in the presence and absence of DAFP-1. If given the opportunity to present, I strictly plan to have my own results to present by February.