Chaperone-Assisted Protein Folding
Skp is an E. coli chaperone initially identified for its role in outer membrane protein biogenesis. Recent work has shown that co-expression of Skp improves periplasmic recombinant protein expression, and Skp is critical for the transport and folding of some bacterial virulence factors, including IscA of Shigella. To elucidate the mechanism by which Skp acts to help fold soluble proteins within the periplasm, wild type and mutant Skp variants will be characterized both in vitro and in vivo for their effects on scFv aggregation and folding. Additionally, Skp's effect on scFv folding will be explored in the context of other periplasmic chaperones as well as molecular crowding. Our results will be used to guide engineering of scFv-specific, ATP-independent chaperones to maximize expression of these valuable protein products. Our results may also help shed light on novel methods to block Skp interaction with important bacterial virulence factors.