Undergraduate Research Initiative – Blog 4
Over the past few months, Dr. Nigel Yarlett and I have been working on characterizing the substrate specificity of Spermidine:spermine N1 – Acetyltransferase (SSAT). In this time we have isolated and expressed the C. parvum SSAT for kinetic studies in order to classify the enzyme either as a true SSAT or a general N-acetyltransferase (GNAT). Previous research has shown that the polyamines: putrescine, spermidine, and spermine have all been found to be essential for the growth, development, and metabolism of C. parvum. SSAT is a major regulator for polyamine synthesis, particularly spermine, and without it the parasite will surely die. Applying chemical kinetic studies allowed us to gauge and study the rates of chemical processes. We can also investigate how different experimental conditions (different substrates) can influence the speed of a chemical reaction and then construct mathematical models, in the form of graphs, describing this chemical reaction based on how much of a certain substrate and varying concentrations are reacting. Initial results indicate that spermine is the preferred substrate for SSAT with minor activity being demonstrated for spermidine, histone 2A, para-amino benzoic acid (PABA), and dapsone. This information was collected through both kinectic analysis of spermine, spermidine, and histone 2A and high–performance liquid chromatography analysis (HPLC) using PABA and dapsone. Upon completion of our kinetic studies and HPLC analysis we may be able to confirm the true substrate specificity for SSAT. If spermine is shown to have the highest activity out of all the substrates tested we can then exploit these biochemical characteristic for rational drug design. This project was very beneficial for me because it has allowed me to use and understand advanced techniques in the science field. My future goal for this project is to synthesize a polyamine analogue structurally similar to spermine. Considering SSAT is substrate specific for spermine, this analogue may be able to act as inhibitor for SSAT, prohibiting parasitic growth and promoting death of C. parvum. These findings will generate a manuscript and will be published in the Journal of Molecular and Biochemical Parasitology. I am glad that I was able to receive so much support from the undergraduate research initiative team in my pursuit of greater knowledge of scientific research. I have learned so many different techniques, how they work, and how I can analyze and interpret raw data. My mentor, Dr. Nigel Yarlett, was definitely inspiring and very supportive in times of frustrations and failures. Overall, I truly enjoyed this experience and thank Dr. Nigel Yarlett and the rest of Haskins Laboratories for this experience.