Unfortunately, due to illness, I have been unable to make it to the lab for 5 days. Consequently, I am a little behind on my project and have not been able to update you all on how my proteins are doing. Whilst lying at home with a fever, I thought of myself as a mother to these poor proteins; without me they don’t stand a chance! The unlucky things have been stuck in the freezer for far too long and today was my chance to transform them so they can be expressed! I actually felt guilty for taking time off and leaving my proteins to fend for themselves. I know… I must have been deliriously ill!
Luckily, I am now reasonably well enough to start work and on my first day back my supervisor showed me the results to the first NMR. Apparently it is the best result we have had yet for the C2 domain of KIBRA. However, we still got 20 peaks less than expected. When I was first told this I didn’t really know what any of it meant, but after some extensive research about NMR on Wikipedia, I began to understand a little better. Basically, for every amino acid in my protein, there should be a peak on the graph. The distance and angles between the peaks can then help us to determine the structure of the protein. In regards to this, we are re-doing our NMR but this time using a buffer with a slightly lower pH to see if this will make a difference.
Meanwhile, I’ve been repeating the entire process of making proteins but for the HACE1 protein instead of KIBRA. Remember I said I was now working on a protein involved in cancer? Well that’s HACE1! Anyway, I’m working on getting my pretty KIBRA protein gels uploaded soon!