The Shoubridge laboratory at the McGill University in Montreal has a long-standing interest in the identification and characterization of genetic defects associated with mitochondrial oxidative phosphorylation deficiencies in both adults and children.
The laboratory’s focus lies in deciphering the function of nuclear encoded mitochondrial proteins and their role in mitochondrial disease pathology at a molecular level.
More recently, we have been using BioID, a proximity biotinylation assay, to create a mitochondrial interaction network. Using this assay, we are studying interfaces between mitochondria and other cellular compartments, especially the endoplasmic reticulum, as well as the functional context of newly identified proteins in mitochondrial RNA granules, the hubs for posttranscriptional regulation of mitochondrial gene expression and the birthplace of mitochondrial ribosomes.
To better comprehend the role of individual functional networks within mitochondria and between mitochondria and the remainder of the cell, we established a mitochondrial interactome using a proximity biotinylation assay, BioID.
BioID uses a promiscuous biotin ligase covalently fused to a protein of interest (bait), which subsequently biotinylates proteins in the vicinity of the bait. Using BioID we can identify novel mitochondrial proteins, determine the localization of an individual protein within the mitochondrion as well as within a functional network, and identify novel roles for proteins both within and outside the mitochondria. In addition, we are investigating how individual interactomes change when mutant protein variants are present, yielding insight into possible pathogenic mechanisms and potential treatment targets. Hana is the specialist when it comes to Bio-ID.