Dear all,
We have a Pro Gradu project working on human MSR3, a protein probably involved in protection against oxidative stress in the secretory pathway. We have cloned the enzyme and shown that we can get soluble expression of the mature protein when recombinantly expressed in E.coli. MSR3 has an unusual C-terminal motif that while not matching the PROSITE motif probably acts to retain the protein in the ER. If MSR3 is, as predicted from the sequence, a methionine sulphoxide reductase then it must interact with a thioredoxin-like protein to complete its catalytic cycle; which of the ER-resident thioredoxin-superfamily members this might be is unknown.
The work will involve:
Purification of recombinantly expressed MSR and analysis of its enzymatic activity
Localization of MSR by immunofluorescence in COS7 and HeLa cells and examination of the effect of the C-terminal motif on localization (requires plasmids to be made by PCR-cloning and mutagenesis)
In vivo protein-protein interaction determination by bimolecular fluorescence complementation, screening for MSR3 interactions with thioredoxin-superfamily members and, time allowing, for the effects of oxidative stress on MSR3-substrate interactions.
Please contact me if you are interested.
Best wishes,
Lloyd