1. 논문 내용과 의미를 설명해 주세요.
Protein oligomerization is an important molecular mechanism in many biological processes that control numerous cellular events. We investigated the property of cryptochrome2 (CRY2) from Arabidopsis thaliana by examining how fusion proteins or tags to influence the efficiency of CRY2 clustering. We found that quaternary structures of tagged proteins, as well as, their conjugation site influenced the efficiency of light-induced CRY2 clustering. This provides valuable information that should be carefully considered prior to tagging and guidelines for designing new optogenetic tools with CRY2. Notably by simply tagging CRY2 with short peptide (CRY2clust) improved CRY2 clustering and showed a propensity for clustering similar to that of native CRY2. We hypothesized that C-terminal extension of CRY2clust may extend existing region for homo-oligomerization and promote its clustering by facilitating hydrophobic interactions. However, further studies are needed to elucidate the mechanism responsible for the enhanced clustering by CRY2clust. The even dispersion of clusters within cells including the nucleus and enhance light sensitivity should widen the scope of CRY2clust application. In fact, application of CRY2clust to the already developed optogenetic system (OptoSTIM1, OptoTRKB and OptoFGFR1) dramatically increase their activity. We expect our CRY2clust will expand the versatility of the optogenetic clustering toolbox and enable researchers to select the precise tool for their experimental purposes.
2. 연구과정에서 있었던 에피소드를 소개해 주세요.
I first started this project in my 2nd year of MS and Ph. D. integrative program. Our initial goal was to conjugate different fluorescent proteins and demonstrate the versatile use of CRY2 for different experimental uses, as we described in our paper. However, generation of CRY2 was not intended. While generating numerous plasmids with different fluorescent proteins, we accidentally found a short peptide could enhance the innate property of CRY2 and story moved on from there. At first, we did not know how to answer this phenomenon. Now thinking back, although we did not intend to create CRY2clust, the process of understanding CRY2clust helped us to further characterize unveiled property of CRY2.
3. 연구를 통해 얻은 지혜를 후배들에게 들려주세요.
From my experiences, I think learning how to operate certain experiments and writing skills are fundamental elements that all graduate students learn throughout their course. Learning how to logically design experiments and to overcome persistent upcoming problems are important tactics learned through research. Also unlike what we all learned through college, studying materials on oneself was not fulfilled. Besides studying and securitizing biological questions, the ability to discuss with colleagues is also one of the important criteria in research. I am still in the middle of learning those processes and hopefully, will earn more insights through further research experiences.
4. 나는 왜 생명과학자가 되었는가?
After graduating pre-medical school, I was in the middle of the road whether I will continue my med school program or learn more biological science for research. I was longing to understand and investigate veiled biological questions that changed my course to become a biologist.
5. 다른 하고 싶은 이야기
The co-author of this paper, Dr. Hyerim Park helped since I started my graduate school. She guided me through this research that we were able to pull this research for better outcomes. I would like to especially thank her for her effects and sincerity.