title: Role of lysines in ion selectivity of bacterial outer membrane porins.
abstract: The epsilon-amino groups of available lysine residues of the OmpC, OmpF and PhoE porin proteins of Escherichia coli and of the protein P porin of Pseudomonas aeruginosa, were modified by the bulky reagent trinitrobenzenesulphonic acid. Approximately 78% of the lysines of the anion-selective protein P and PhoE porins were modified whereas only 40-50% of the lysines of the cation selective OmpF and OmpC porins were altered. After modification, the three E. coli porins had very similar high selectivities for cations over anions, in contrast to the native porins which varied 86-fold in ion selectivity. Despite the large size of the trinitrophenyl group attached to modified lysines (i.e., a disc of approx. 0.86 nm diameter X 0.36 nm high) relative to the reported size of the constrictions of the E. coli porins (1.0-1.2 nm diameter), only the anion-selective PhoE porin was substantially blocked after trinitrophenylation. The protein P porin channel was relatively unaffected by trinitrophenylation, in contrast to previous data showing dramatic effects of acetylation of lysines on protein P conductance and selectivity. This favoured a model in which the critical lysines involved in anion binding by protein P were present in a constriction of the channel that was too small for trinitrobenzenesulphonic acid to enter. Overall, the data suggest that both the number and relative position of charged lysines are major determinants of ion selectivity.
mesh_terms: D000838:Anions; D001425:Bacterial Outer Membrane Proteins / Q000502:physiology*; D002412:Cations; D002463:Cell Membrane Permeability; D055598:Chemical Phenomena; D002621:Chemistry; D002712:Chlorides / Q000378:metabolism; D004553:Electric Conductivity; D004926:Escherichia coli / Q000032:analysis; D007473:Ion Channels / Q000502:physiology*; D008239:Lysine; D008564:Membrane Potentials; D018272:Porins; D011188:Potassium / Q000378:metabolism; D011550:Pseudomonas aeruginosa; D013329:Structure-Activity Relationship; D014302:Trinitrobenzenesulfonic Acid
chemical_list: D000838:Anions; D001425:Bacterial Outer Membrane Proteins; D002412:Cations; D002712:Chlorides; D007473:Ion Channels; D018272:Porins; D014302:Trinitrobenzenesulfonic Acid; D008239:Lysine; D011188:Potassium