My Protein Didn't Fold and Neither Did I
When Gaurav Ghag realized that he had replicated a calculation error in every experiment during four years of his graduate research, he initially thought that his career had unraveled with his protein.
In 2016, I was a fourth-year graduate student at the University of Southern Mississippi. I studied the structure of granulins, which are cysteine rich proteins implicated in neurodegenerative diseases. To verify protein folding, I conducted an assay so often that I created a templated, ready-to-use protocol for it.
As I inched towards the end of graduate school, I was excited. I had fantastic data on my folded protein, and a high impact journal publication seemed within reach.
One day, I tested an alternative method to assess granulin folding, and weirdly, it indicated that the protein was unfolded. Perplexed, my advisor, Vijayaraghavan Rangachari, and I walked through every step of the experiment until it struck me: I had made a decimal point calculation error in my templated protocol! I realized with dismay that I had replicated this mistake in every experiment. I had been working with unfolded protein all along.
Shocked, I exited my advisor’s office, poured myself some coffee, and just sat outside for hours in disbelief and disappointment. I had no time to start a new project or to repeat my experiments. What was I going to do?
Once I calmed down the next day, I decided that it wasn't over. I still had highly reproducible (unfolded) protein data that might be worth something. With the support of my advisor, I managed to publish the data.1 It didn't end up being the high impact factor paper that I dreamed of, but we managed to put a positive spin on a bad situation.
I view that incident as the biggest learning moment of my life. First, the devil is in the details, so I always have someone else double check critical work now. Second, I remind myself that mistakes happen, but resilience helps overcome them.
Although I have moved on since, I often remember a thought from that day: "Cysteines, thou art heartless beasts!"
This interview has been edited for length and clarity.
- Ghag G, et al. Protein Eng Des Sel. 2016;29(5):177-186.