I want to express a few proteins in yeast and mammalian cells which should be translocated immediately to the mitochondria upon expression. Any vector or signal sequence information would be helpful.
One idea is, use the mt-specific signal. A protein I have cloned, cardiolipin synthase, is localized EXCLUSIVELY in the inner mt membrane. At4g04870 has 38AA signal peptide, which you can use for yeast expression (look up my FEBS paper and PhD thesis). After this sequence, you could clone-in regular FLAG sequence to ensure you can remove the targeting sequence.
GL and let me know how it worked!
I heard it from my colleague and found the following article that may be useful to you.
Gerard G.M. D'Souza, Sarathi V. Boddapati, Volkmar Weissig
Mitochondrial leader sequence-plasmid DNA conjugates delivered into mammalian cells by DQAsomes co-localize with mitochondria.
Mitochondrion Volume 5, Issue 5, October 2005, Pages 352–358
I think if you can replace the leader sequence of the targeting protein with a leader sequence of a known mitochondrial matrix protein while cloning, then it can lead protein directly to mitochondrial matrix and you may get its expression in that particular compartment.
Good luck with your experiments !
Agreed- if you add a signal sequence you can get it to go directly to the mito. For a matrix-targeted protein see my paper attached
Del Gaizo V, Payne RM. Mol Ther. 2003 Jun;7(6):720-30.PMID: 12788645.
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