Metabolic Research Unit - MRU
Protein function can be elucidated from the study of protein-protein interactions.
The target’s interacting partners are screened using a Yeast 2- hybrid screening technique. Other techniques such as BRET and BIAcore are utilised to confirm/disprove hypotheses and to further analyse the nature of the interaction.
Yeast 2-hybrid
Yeast 2-hybrid screening is an in vivo genetic yeast-based system that identifies candidate protein ‘partners’ that interact with the protein encoded for by the candidate gene.
BIAcore technology
BIAcore Technology is used to analyse interactions between two or more molecules in real time.
Association/dissociation can be monitored in real time.
It is an excellent tool for studying weak to moderate /strong interactions.
BIAcore can be applied to study interactions such as:
BRET - Bioluminescence Resonance Energy Transfer
FRET - Fluorescence Resonance Energy Transfer
Both BRET and FRET are excellent tools for studying protein interactions as they occur in intact live cells.
BRET is based on energy transfer from a bioluminescent donor to a fluorescent acceptor.
The ratio of green to blue light increases as the two proteins are brought into close proximity, and is measured as a BRET signal.
Other applications for BRET include:
in Yeast-2 hybrid screens.
