Abstract
The 'seventeen kilodalton protein' Skp confers transient solubility on outer membrane proteins during biogenesis in Gram-negative bacteria. Here we report a first biophysical characterization of this chaperone itself, which also possesses biotechnological potential in the production of recombinant proteins. Using cross-linking and gel filtration methods, we found that Skp forms a stable homo-trimer in solution. Following thermal denaturation, monitored by CD spectroscopy, this chaperone refolds with high efficiency but exhibits a pronounced hysteresis between the un- and refolding transitions. Using the recombinant protein equipped with the Strep-tag II at its N-terminus, suitable crystallization conditions for Skp were found. A first data set was collected to 2.60 Å resolution.
Original language | English |
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Pages (from-to) | 137-143 |
Number of pages | 7 |
Journal | Biological Chemistry |
Volume | 385 |
Issue number | 2 |
DOIs | |
State | Published - Feb 2004 |
Keywords
- Circular dichroism
- Cross-linking
- Outer membrane protein
- Protein folding
- Strep-tag
- Thermal denaturation