Sample interview questions: Can you provide examples of how you have used proteomics to study protein-protein interactions?
Sample answer:
- Affinity purification-mass spectrometry (AP-MS): I have utilized AP-MS to identify proteins that interact with a protein of interest. This technique involves immobilizing the protein of interest on a solid support and then incubating it with a cell lysate or tissue extract. The interacting proteins are then eluted from the solid support and identified by mass spectrometry.
- Co-immunoprecipitation (Co-IP): I have also used Co-IP to identify protein-protein interactions. This technique involves immunoprecipitating a protein of interest and then analyzing the immunoprecipitated complex by mass spectrometry. The proteins that are co-immunoprecipitated with the protein of interest are likely to be interacting partners.
- Proximity labeling: I have employed proximity labeling techniques, such as BioID and APEX, to identify proteins that are in close proximity to a protein of interest. These techniques involve fusing a promiscuous biotin ligase or peroxidase enzyme to the protein of i… Read full answer
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