Shiana: Promising Beginnings

This last week and couple of days have been more than I knew to expect. I was surprised at how comfortable I felt with the group of students I was with within the first day of our internship, and then again pleasantly surprised when the comment I made voicing as much was agreed with. Training in a classroom together with Nadja that first day for some four hours did the trick, all right. Despite my existing background in laboratory techniques, Nadja, Jeff, Rachel, and the other students all helped make the review of knowledge fun for me. Much of our days were spent smiling or laughing in between our mini-labs and lectures, both at each other and at side jokes. The presenters that we had the chance to listen to this week were so enthused and animated about their own work that it was hard for me to not be drawn into their projects.

Having been assigned to a lab in the College of Pharmacy, I was required to take an extra toxicology course this second week. I readily appreciated the chance for this additional background before I was fully immersed in my lab work; for a surety, it helped me to better understand the research papers that I had to read for my lab. The teachers and presenters during this segment of learning, which this week included again Marti and Katy, were very willing and patient about teaching us what they thought we could benefit from. Their support and the friendliness of the BIO5 staff I have met so far has definitely made it easier to ask for help and interact with them.

Onward ho to lab life--the exciting part! My mentor's name is Ruiyu Xie (but I call her Rae, for short) and she has been working at the College of Pharmacy for at least three years. She is very easy to approach with questions and explains concepts to me I have not learned yet patiently. Through the past four days we've talked about everything from our backgrounds, families, and friends to even troublesome laptop problems. Dr. Monks was a delight to conversate with when I met with him on Tuesday, as well. He first came from the UK, hence his accent, and came to the U.S. to continue his studies. I also met Rae's best friend who works under Dr. Lau, Chi Lin, who I discovered knew my parents! I had no idea. Not only that, but a former student of my high school and a friend of my older sister's, is also volunteering in Dr. Lau's lab this summer. Evidently, it's a small world. I have also made friends with so many other undergraduate and graduate students that I look forward to seeing them every day around the lab.

Research-wise, I will be working with a mitochondrion-specific peroxidase called peroxiredoxin III (Prx III). Past research suggests that the enzyme Prx III is a critical regulator in the redox stage of a cell. Reactive oxygen species (ROS) must be reduced ultimately to compounds that consist of mainly water so that they do not harm the cell. Catalase, glutathione peroxidase (GPx), and Prx can all reduce a ROS called hydrogen peroxide, but the mitochondrion lacks catalase and must import glutathione (GSH). The only enzyme that it inherently has to perform this job is Prx; however, specifically Prx III in mitochondria. Past research has discovered that there is about 30 times more Prx III than GPx1 in the mitochondria. This seems to indicate that Prx III may have more roles than the regulation of apoptosis in the mitochondria.

In order to further determine the function of Prx III, I will render Prx III unable to function in a live cell and observe what cell processes no longer proceed normally because of its 'absence.' To disable Prx III, I will mutate it by replacing two specific amino acids in it called cysteine (Cys) 108 and Cys 229 with another amino acid called serine (Ser). These two particlar Cys amino acids were chosen because they have been found in past studies to form the disulfide bond necessary for Prx III to function. Ser was chosen to replace Cys because it is the most similar in structure to Cys and it will not change the three dimensional structure of Prx III.

So far, Rae and I have sent in a viral plasmid to be sequenced (for which reasons will be saved for another entry) and must wait until the results are returned in order for us to proceed with the next step of our experiment. Though our plasmid was sent in two days ago and were expected to be finished sequenced before today, we should receive them for sure tomorrow. Then, the real lab work begins. Meanwhile, we have designed two mutagenesis primers that we ordered today--a process that included locating and then translating the Prx III's nucleotides into amino acids, locating the specific amino acids of interest in terms of nucleotides, figuring calculations, and comparing primer possibilities to find the optimal ones. The completion of this project will be beneficial to the research of a great many people working in the same field, and I am more than ready to put my eager energy and time into it.