Products/Services Used | Details | Operation |
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Proteins, Expression, Isolation and Analysis | Concentrated Skp was denatured with 6 M Gdm/HCl, applied to Ni2+ beads (Genscript), and eluted with 300 mM imidazole. | Get A Quote |
The biogenesis of integral outer-membrane proteins (OMPs) in Gram-negative bacteria requires molecular chaperones that prevent the aggregation of OMP polypeptides in the aqueous periplasmic space. How these energy-independent chaperones interact with their substrates is not well understood. We have used high-resolution NMR spectroscopy to examine the conformation and dynamics of the Escherichia coli periplasmic chaperone Skp and two of its complexes with OMPs. The Skp trimer constitutes a flexible architectural scaffold that becomes more rigid upon substrate binding. The OMP substrates populate a dynamic conformational ensemble with structural interconversion rates on the submillisecond timescale. The global li... More