Effect of various mild surfactants on the reassembly of an oligomeric integral membrane protein OmpF porin.
Reassembly of OmpF porin from its denatured monomer into the sodium dodecyl sulfate-resistant species was investigated by using 27 kinds of mild surfactants. Polyethyleneoxide-type surfactants with a hydrophilic-lipophilic balance value of 10.8-14.6 induced the trimerization of denatured OmpF porin. Dimerization and trimerization were induced by non-polyethyleneoxide-type mild surfactants that are generally used for membrane protein solubilization. The dependence of surfactant concentrations on reassembly was estimated to obtain a minimal concentration required for the reassembly of the protein. Extensive reassembly (to approximately 85% yield) into dimer (a putative assembly intermediate) was observed at a protein concentration of 0.05 mg/ml in 7 mg/ml n-octyl-beta-D-glucopyranoside and 1 mg/ml sodium dodecyl sulfate. This condition will be useful for the studies of the dimer and dimerization of OmpF porin. The role of mixed micelle system on the protein renaturation was discussed.
Watanabe
Yasushi
Y
National Food Research Institute, Tsukuba, Ibaraki, Japan. yasuw@nfri.affrc.go.jp
D002850:Chromatography, Gel; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry; D008823:Micelles; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000032:analysis; D018272:Porins / Q000737:chemistry*; D017510:Protein Folding; D013501:Surface-Active Agents / Q000737:chemistry*
D029968:Escherichia coli Proteins; D008823:Micelles; C109237:OmpF protein; D018272:Porins; D013501:Surface-Active Agents; D011092:Polyethylene Glycols