Diogo’s update on his last sprint of his PhD

– Could you remind us of what is your project about?

My work is about making a time-resolved X-Ray crystallography experiment with membrane proteins.

In order to do this, I have to purify enormous protein quantities and then obtain crystals. These crystals are then measured with X-Rays and we acquire many 3-D protein “snapshots”, we then put them together following a time-line, in the end obtaining a “movie” of the protein in action.

Could you give us a brief update on your project?

My project like many others took twists and turns, and was hampered by the recent pandemic that affected all of us worldwide. Now however, I am reaching the final sprint of the project; I have purified and obtained crystals of the mutant version of my current  membrane protein target; I have validated that it is in the state that we need it to be; I have troubleshot some additional problems that popped up during the protein production; and am now producing large amounts of the protein for the time-resolved experiments.

I am now producing daily about 10 L worth of bacteria culture that produce my protein. Due to the current restrictions related to the pandemic I can’t work every day to produce this protein, but am getting some nice chunks of time to do it efficiently. In the near future I also will have access to large scale fermenters, very similar to the fermenters that are used to produce beer from yeast. These are large tanks that constantly control the temperature, oxygen levels, pH (acidity), and some times nutrient levels of the microorganism of interest, making large scale protein production much more efficient.

               I now need to work a lot in the lab for the last sprint of my project, and with a bit of luck, there won’t be more unforeseen obstacles and I will have an amazing experiment on the first attempt. And if new problems arise throughout the way, well, that’s what usually happens and we will work out a new way around the problem!

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