The periplasmic skp chaperone has been characterized primarily for its role in outer membrane protein (OMP) biogenesis, in which the jellyfish-like trimeric protein encapsulates partially folded OMPs until delivery to the outer membrane. However, skp is being increasingly recognized as a mediator of soluble protein folding; skp co-expression increases the active yields of many recombinant proteins and bacterial virulence factors. The molecular mechanisms governing soluble protein recognition by skp are currently unknown. Using a panel of single-chain variable fragments (scFvs), we performed in vitro folding and aggregation assays in the presence or absence of skp. Kinetic data for skp-sensitive scFvs were fitted globally to a model whereby skp acts to reduce aggregation by sequestering partially folded intermediates. Folding intermediates were further confirmed by equilibrium denaturation. These results will aid future engineering projects aimed at achieving high-level recombinant protein yields in E. coli and identify novel approaches to block bacterial virulence.

<i>Escherichia coli</i> skp chaperone sequesters kinetically trapped intermediates of soluble proteins to reduce aggregation

243rd National Meeting (2012)

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Escherichia coli skp chaperone sequesters kinetically trapped intermediates of soluble proteins to reduce aggregation