Difference between revisions of "Part:BBa K4244049"

Revision as of 09:35, 9 October 2022

flip-amilCP (MMP-9 inducible)

This flip-amilCP is designed in such a way that it will only regain its colour when it is cleaved by MMP-9. Two inserts have been made, the E5 and K5 domain. These two domains dimerize and disrupt amilCP structure. MMP-9 recognizes the cleavage site and cuts off the K5 domain, restoring Ultramarine function.

The flip design is based off the following paper: Zhang, Q., Schepis, A., Huang, H., Yang, J., Ma, W., Torra, J., Zhang, S. Q., Yang, L., Wu, H., Nonell, S., Dong, Z., Kornberg, T. B., Coughlin, S. R., & Shu, X. (2019). Designing a Green Fluorogenic Protease Reporter by Flipping a Beta Strand of GFP for Imaging Apoptosis in Animals. Journal of the American Chemical Society, 141(11), 4526–4530. https://doi.org/10.1021/JACS.8B13042/SUPPL_FILE/JA8B13042_SI_002.PDF"

The structure of flip-amilCP was predicted using alphafold as shown below.

The left shows uncleaved flip-amilCP, the middle the cleaved, active flip-amilCP, and the right regular amilCP

Sequence and Features