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Darren Boehning

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Rapid and transient palmitoylation of the tyrosine kinase Lck mediates Fas signaling.
Sep 23, 2015   Proceedings of the National Academy of Sciences of the United States of America
Akimzhanov AM, Boehning D


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The Long Road To Publication

Sep 30, 2015

Have you ever started reading a paper and said to yourself that you could have done this work in a couple of months? Looking at our recent paper, with four relatively small figures and no animal work, you could easily come to this conclusion. However, this paper was a long time in the making. Actually, I could not even remember precisely when we came up with the idea. I dropped into the office of co-author Askar Akimzhanov and asked him if he remembered. We were studying the activation of a T-cell kinase called Lck. It is easy to measure the activity of this kinase by Western blotting because it is phosphorylated in the active state. However, when we stimulated the cells the phosphorylated form of the kinase did not increase as expected. Although I don’t remember saying this, Askar told me that I suggested he look in the detergent insoluble pellets. Sure enough, that is where our missing active Lck was located. This was the initial piece of data which ultimately led us to hypothesize that Lck is rapidly palmitoylated in response to a given stimulus.  We both estimated this was around eight years ago. For reference, I have been a faculty member for a little over ten years. Askar was a post-doc at the time, and is now faculty.


Why did this paper take so long? It was not for a lack of effort. The main hurdle turned out to be technical. In brief, there simply was not a sensitive enough assay to detect the palmitoylation of what is likely a small sub-pool of the total Lck protein. How we overcame that technical limitation is almost laughably illustrated as a simplistic cartoon in figure two of the paper. The details are unimportant for discussion here, other than to state that the majority of our time was spent figuring out how to do this assay. Once we could do it reproducibly, the rest was relatively easy, and some of our results were surprising. For example, not only was Lck palmitoylated with incredibly fast kinetics, it was de-palmitoylated almost as fast. We now have the framework for studying the palmitoylation of many other substrates which will likely occupy at least another ten years of work.


Certainly the road to completion for this project was extremely difficult, especially for Askar. I had essentially given him an ultimatum at least twice that if he didn’t get it working in a few months, we would have to scrap the project. To add insult to injury, the grant he was funded on went into no-cost extension, and because of this and visa issues he had to return to his native Kazakhstan for half a year before funding was restored on the project.  In a scientific fairy tale, Askar fell in love with his wife-to-be while in Kazakhstan. Sometimes in science, the struggles have a silver lining.






Copyright: © 2013 Darren Boehning. The above content is licensed under the Creative Commons Attribution License (CC-BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.  
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