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 A resolution.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Circular Dichroism
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Crystallization
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Crystallography, X-Ray
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DNA-Binding Proteins / chemistry*
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DNA-Binding Proteins / genetics
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Escherichia coli / metabolism
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Escherichia coli Proteins / chemistry*
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Escherichia coli Proteins / genetics
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Hot Temperature
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Molecular Chaperones / chemistry*
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Molecular Chaperones / genetics
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Molecular Sequence Data
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Periplasm / metabolism
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Periplasmic Proteins / chemistry*
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Periplasmic Proteins / genetics
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Protein Denaturation
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Protein Renaturation
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Protein Structure, Quaternary
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Protein Structure, Secondary
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
Substances
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DNA-Binding Proteins
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Escherichia coli Proteins
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Molecular Chaperones
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Periplasmic Proteins
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Recombinant Proteins
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Skp protein, E coli