To study the molecular mechanisms of human MICAL1, we first produced this protein in insect cells using a baculoviral expression system. With single-particle cryoelectron microscopy, we determined the overall architecture of MICAL1 and solved the structure in very fine detail. We were the first ones to see the full-length MICAL structure, which helped us understand how it works. The overall architecture of MICAL1 revealed that a C-terminal domain folds over the catalytic site, restricting substrate access. This intramolecular interaction between the C-terminal and N-terminal catalytic domains appears to impose MICAL1 autoinhibition, which we confirmed through biochemical and biophysical studies.
We also tried to understand how MICAL works with other proteins called plexin receptors. Unfortunately, we were not able to detect any binding between them. So, we tried using another protein called Rab8, which we found in previous studies. Testing Rab8 and MICAL together, we observed direct binding. However, we have not yet determined the structure of the complex due to difficulties with plexin. Nevertheless, this project is ongoing, and we expect to determine the structure of the complex soon.
Another goal of the project was personal growth, and I believe this goal was fully addressed. In particular, I significantly improved my leadership and management skills by attending four workshops focused on project management. My leadership skills were further developed by supervising two undergraduate students.
The findings of this research will be published soon, and we expect to submit two manuscript by the end of this year. I presented the results through conferences and a number of invited talks. Our research was also popularized through newspaper interviews.
