Pore-forming activity of OmpA protein of Escherichia coli.
Escherichia coli outer membrane protein OmpA was purified to homogeneity, as a monomer, from a K12 derivative deficient in both OmpF and OmpC porins. When proteoliposomes reconstituted from the purified OmpA, phospholipids, and lithium dodecyl sulfate were tested for permeability to small molecules by osmotic swelling, it was found that OmpA produced apparently nonspecific diffusion channels that allowed the penetration of various solutes. The pore-forming activity was destroyed by the heat denaturation of the OmpA protein, and the use of an OmpA-deficient mutant showed that the activity was not caused by copurifying contaminants. The size of the OmpA channel, estimated by comparison of diffusion rates of solutes of different sizes, was rather similar to that of E. coli OmpF and OmpC porins, i.e. about 1 nm in diameter. The rate of penetration of L-arabinose caused by a given amount of OmpA protein, however, was about a hundredfold lower than the rate produced by the same amount of E. coli OmpF porin. The addition of large amounts of lithium dodecyl sulfate to the reconstitution mixture increased the permeability through the OmpA channel, apparently by facilitating the correct insertion of OmpA into the bilayer.
Sugawara
E
E
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Nikaido
H
H
D001089:Arabinose / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D004058:Diffusion; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D007473:Ion Channels; D010743:Phospholipids / Q000378:metabolism; D011489:Protein Denaturation; D011510:Proteolipids; D012967:Sodium Dodecyl Sulfate / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D007473:Ion Channels; D010743:Phospholipids; D011510:Proteolipids; C020485:proteoliposomes; D012967:Sodium Dodecyl Sulfate; D001089:Arabinose; C028913:dodecyl sulfate