Team:Hong Kong HKUST/Project/module3

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<h3>Mechanism of MLS</h3>
<h3>Mechanism of MLS</h3>
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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.
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<p>MLS is attached to the N-terminal of the protein in interest. The MLS will be recognized by receptor on outer mitochondrial membrane. The complex of receptor and attached protein will diffuse to contact site, where the outer and inner membrane come into contact. The protein will be translocated across mitochondrial membrane via a protein translocator. The protein will be unfolded and chaperone will pull the protein across the membrane. MLS will be cleaved off afterward. The refolding of protein will be done by chaperone.</p>
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<img src="https://static.igem.org/mediawiki/2013/8/87/MLS_mechanism.png" >
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Revision as of 17:36, 27 September 2013

Protein Trafficking

Overview

In vertebrates, the signal sequence coding region promotes translocation of precursor peptide. 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 glyoxylate enzymes (AceA and AceB proteins, denoted ACE enzymes) should be targeted to mitochondria for their functionality. To do so, we fused ACE enzymes with Mitochondrial Leader Sequence (MLS). We have constructed and qualitatively and quantitatively characterized Mitochondrial Leader Sequence BioBrick BBa_K1119001.

Mechanism of MLS

MLS is attached to the N-terminal of the protein in interest. The MLS will be recognized by receptor on outer mitochondrial membrane. The complex of receptor and attached protein will diffuse to contact site, where the outer and inner membrane come into contact. The protein will be translocated across mitochondrial membrane via a protein translocator. The protein will be unfolded and chaperone will pull the protein across the membrane. MLS will be cleaved off afterward. The refolding of protein will be done by chaperone.

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.

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