Team:Hong Kong HKUST/Project/module3

Overview

In nature, eukaryotic cell mitochondria usually have their protein encoded by gene in nucleus and produced from cytosol. In this process, MLS will act as signal peptide to target the protein into mitochondria.
In our module, we will construct the MLS BioBrick K1119001, and characterize it quantitatively.

Mechanism of MLS

MLS is attached to the N-terminal of enzyme, and bind to the receptor protein on mitochondrial membrane, and diffuse to contact site where inner and outer membrane fuse, then bring the ACE enzyme into mitochondria. Afterward, it is be cleaved, leaving the enzyme in mitochondria.

Linkage to project

In our project, we introduced bacterial enzymes to mammalian cell to modify metabolic pathway. However, unlike bacteria, citric acid cycle in mammalian cells is compartmentalized in mitochondria. The ACE proteins should be targeted to mitochondria for their functionality. To do so, we fused ACE enzymes with Mitochondrial Leader Sequence (MLS).

Reference

Alberts, B., Bray, D., Hopkin, K., Johnson, A., Lewis, J., Raff, M., Roberts, K., & Walter, P. (2010). Essential cell biology. (3rd ed., p. 505). UK: Garland Science.