Team:Hong Kong HKUST/module3backup

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 [link], and characterize it quantitatively[link].

Mechanism of MLS

MLS is attached to the N-terminal of the protein in interest, and be recognized by receptor in the outer mitochondria membrane. The complex of receptor and attached protein diffuse to a contact site, where protein is translocated across both the outer and inner membrane by a protein translocator. The MLS will be cleaved off by signal peptidase inside the mitochondria afterward. Chaperone proteins will help to pull the protein across the membrane and help the refolding of protein.

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.