Protein degradation is vital for all living organisms and needs to be precisely regulated. In bacteria, it is mainly performed by ATP-dependent proteases. The Lon AAA+ protease is highly conserved; however, the molecular mechanisms regulating Lon activity are not fully understood and little is known about its cofactors and adaptors. Previous work in our lab identified the heat shock protein LarA as a novel Lon substrate and regulator of Lon in Caulobacter crescentus. LarA accumulates at the onset of heat stress and allosterically activates Lon via its C-terminus. My PhD project is aimed at a better mechanistic understanding of the activator-mediated regulation of Lon. LarA was identified thanks to multiple proteomic-based approaches. We are currently working on the identification of other Lon substrates and adaptors. We aim to short-list our protein candidates from the proteomics' dataset by combining a protein-protein interactions (PPIs) prediction tool. We will use the AlphaPulldown python package published Yu et al. in November 2022 (DOI: 10.1093/bioinformatics/btac749). This package uses the artificial intelligence AlphaFold2 Multimer structure prediction tool to screen potential interactors of a protein of interest according to the predicted structures. We will screen Caulobacter Lon against the whole Caulobacter proteome to pull down potential Lon interactors.