Research
Developing tools to measurE proteostasis
Classically single metastable or unstable proteins (aggregation prone proteins) are used to check if cells can fold these test proteins under different conditions. However, each protein (and each mutant) has its own preferred chaperone interactors (nodes) making it difficult to probe the functionality of all the nodes of proteostasis using a single misfolding mutant.
We device assays using a pool of mutant proteins to assay the functionality of multiple nodes of proteostasis network (PN) in parallel. We take advantage of deep-mutational scanning with the help of in-house high throughput sequencing facility.
Using Adaptive Laboratory Evolution to identify new members of PN
Laboratory Adaptive Evolution is a powerful technique that allows us to evolve E. coli or yeast strains with better proteostasis. We use these strains to understand the pathways cells rely on to reinforce proteostasis during chronic misfolding stress.
While not limited by techniques, we use a combination of different omics techniques, protein biochemistry, protein biophysics and in vitro reconstitution assays to identify new members and components of PN. Our focus is to identify different metabolites, their pathways and cellular signaling processes that connect the concentration of the metabolites with problems in cellular proteostasis.
Delineating the effect of proteostasis on genome evolution
We have preliminary evidence to suggest that even single mutations that lead to protein misfolding may destabilize the whole genome. We are trying to understand the process in order to predict the course of evolution given a misfolding event in the proteome.
We further wish to use the same system to identify new components of proteostasis network and check if metabolism affects proteostasis in eukaryotes too.