Determination of ion permeability through the channels made of porins from the outer membrane of Salmonella typhimurium in lipid bilayer membranes.
The three types of porin (matrix-proteins) from Salmonella typhimurium with molecular weights of 38,000, 39,000 and 40,000 were reconstituted with lipid bilayer membranes either as a trimer or as an oligomer (complex I). The specific conductance of the membranes increased several orders of magnitude after the addition of the porins into the aqueous phase bathing the membranes. A linear relationship between protein concentration in the aqueous phase and membrane conductance was found. In the case of lower protein concentrations (10)(-12)M), the conductance increased in a stepwise fashion with a single conductance increment of 2.3 nS in 1 M KC1. For a given salt the conductance increment was found to be largely independent of the particular porin (38 K, 39 K or 40 K) and on the state of aggregation, although porin oligomers showed an up to 10 times smaller conductance increase in macroscopic conductance measurements. The conductance pathway has an ohmic current voltage characteristic and a poor selectivity for different alkali ions. Further information on the structure of the pores formed by the different porins from Salmonella was obtained from the selectivity for various ions. From the permeability of the pore for large ions (Tris+, glucosamine+, Hepes-) a minimum pore diameter of 0.8 nm is estimated. This value is in agreement with the size of the pore as calculated from the conductance data for 1 M KC1 (1.4 nm for a pore length of 7.5 nm). The pore diameter may well account for the sugar permeability which has been found in reconstituted vesicles. The findings reported here are consistent with the assumption that the different porins form large aqueous channels in the lipid bilayer membranes and that the single conductance unit is a trimer. In addition, it is suggested that one trimer contains only one pore rather than a bundle of pores.
D001426:Bacterial Proteins / Q000494:pharmacology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D002951:Citrates / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004553:Electric Conductivity / Q000187:drug effects; D006531:HEPES / Q000378:metabolism; D066298:In Vitro Techniques; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000494:pharmacology*; D012486:Salmonella typhimurium / Q000378:metabolism*; D014325:Tromethamine / Q000378:metabolism
D001426:Bacterial Proteins; D002951:Citrates; D008051:Lipid Bilayers; D008565:Membrane Proteins; D014325:Tromethamine; D006531:HEPES