Last week, I had a committee meeting. In preparation for the meeting, I met with my PI to discuss how to rework my aims, change my hypothesis, and the best way to lay out the “future manuscript” my committee likes to see (i.e., when I acquire data, I will put it into a paper like so). This lead to a very long discussion in which my PI decided that I had too much chemistry in my project (too MUCH chemistry? blasphemy! there is no such thing.) and wanted to try to shift it back to the biochemistry/biophysics side of things.
Now, let’s backtrack a wee bit here. My second and third years of graduate school were encompassed mostly by failed purification attempts of a fragment of a particular protein which binds to the main protein studied by the lab. Let’s call this protein fragment Protein Y. Now, I was not the first person to be assigned the task of purification; rather, a post-doc in the lab and our protein purification lab tech had each sacrificed months of their time in lab attempting to purify Protein Y with no success. I, however, was not aware of the previous failed attempts, and presuming Protein Y to be like any other protein for which I found a purification strategy, forged ahead with elaborate plans of what to do with said protein, including crosslinking and mass spec to identify the region where Protein Y binds to Protein X, and attempts at co-crystallzation, thinking that perhaps Protein Y would stabilize Protein X enough to form crystals.
I then spent two miserable years trying to purify protein Y, with no success at all. I certainly do not consider this time to be a waste… rather, I acquired so much knowledge about protein purification and various ways to purify a protein that I have now become the resident protein purification expert. The chair of biochemistry has informed me that there is quite the demand for biochemist post-docs with expertise in protein purification, so even though the project tanked, I still think it was integral to my future purification success and perhaps even obtaining a future position.
The Protein Y Purification Project never officially closed, but rather, my time was shifted to working on other projects to help get a paper out the door. I, naively perhaps, assumed that since my thesis aims had shifted to reflect new developments in the lab, and a project which interested me much more (more chemistry! more biophysics!), I was off the hook from my dreaded Protein Y purification days.
Until last week, when I met with my PI. He wants me to go back to Protein Y purification, and re-visit the crosslinking and co-crystallization ideas. Nevermind that the lab has now spent 3+ years trying to purify the protein, racking the brains of one post-doc, one protein guru technician, and one very disgruntled graduate student. I moaned. I groaned. And boy, did I complain. And then, I offered up a perfectly acceptable solution for determining the binding location without having to purify the protein. My PI thought about this, and agreed that it would be a good alternative… but really, he’d rather just have me purify the protein. I have no doubt that this has less to do with my own experiments, and more to do with all the things he thinks he could do with a purified protein after I leave… after all, his priority is the future of the lab, and not getting me out the door in a timely fashion.
At my committee meeting, after I presented everything (quite well, according to my PI), the chair of my committee asked what I thought the biggest challenge would be to completing my aims. Without hesitation, I pointed directly to Protein Y as the source of my likely shortcomings. The other members of my committee nodded and agreed, after witnessing two years of my failed attempts. But, never one to be discouraged or accept that perhaps a protein just cannot be purified, my PI immediately jumped up and went through a spiel about how I am a better biochemist now than I was two years ago and how I will certainly have much more success this time.
After just one more week back with Protein Y, I already want to strangle myself. I wonder how many more years I will have to spend on this before my PI accepts that maybe, just maybe, my research is impossible.