Abstract
The in vitro reassembled species of OmpF porin, which was renatured from its denatured monomer using n-octyl-β-D-glucopyranoside, was characterized by low-angle laser light scattering photometry, circular dichroism spectroscopy and synchrotron radiation small-angle X-ray scattering measurements. The light scattering measurement reconfirmed that the reassembled species was the dimer of the protein. Circular dichroism spectra of the reassembled dimer showed a native-like β-structure. A small-angle X-ray scattering measurement indicated that the size of the reassembled dimer was nearly equal to that of the native trimer under the present experimental conditions. In a thermal denaturation experiment followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the reassembled dimer was less stable than the native trimer.
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Abbreviations
- CD:
-
circular dichroism
- GPC:
-
gel permeation chromatography
- MOPS:
-
3-[N-morpholino]propanesulfonic acid
- OG:
-
n-octyl-β-sc d-glucopyranoside
- SAXS:
-
small-angle X-ray scattering
- SDS:
-
sodium dodecyl sulfate
- UV:
-
ultraviolet
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Watanabe, Y., Inoko, Y. Physicochemical Characterization of the Reassembled Dimer of an Integral Membrane Protein OmpF Porin. Protein J 24, 167–174 (2005). https://doi.org/10.1007/s10930-005-7840-7
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DOI: https://doi.org/10.1007/s10930-005-7840-7