Monitoring the mitochondrial localization of mycobacterial proteins.

Mitochondrion apart from being the energy hub of the cell, is also the center of various signaling pathways. During intracellular infection, either bacterial or viral, several pathogen proteins, metabolites, and possibly, lipids interact with the host mitochondria. These interactions allow the pathogens, such as Mtb, to reprogram the host mitochondrial functions, screwing the host immune responses and resulting in the persistence of the bacteria. Therefore, mitochondria offer a critical target organelle for various therapeutic interventions. This chapter deals with methods to demonstrate and establish the mitochondrial localization of Mtb proteins by confocal microscopy and mitochondrial enrichment. Transient transfection of mammalian constructs or infection Mycolicibacterium smegmatis infection could be used to overexpress the candidate Mtb protein in host cells, allowing the study of changes in the mitochondria's composition and function with regard to localization studies, mitochondrial DNA and RNA, proteomics, metabolomics, lipidomics, and bioenergetics.