Effect of point mutations on the in-vitro pore properties of maltoporin, a protein of Escherichia coli outer membrane.
Maltoporin (LamB protein), a protein of Escherichia coli outer membrane forms ionic channels with a selectivity for maltose and maltodextrins (Dargent et al., 1987). The effect of different point mutations on maltoporin pore properties was investigated in vitro with planar bilayers. The mutations belong to three classes in terms of selective maltose transport in vivo: class A (substitution at positions 259 and 382) does not affect maltose transport, class B (position 163 and 245) decreases maltose transport down to 20 to 30%, and class C (position 18) almost completely abolishes selective maltose transport. This in-vitro study reveals that class A does not affect the pore properties in contrast to class B substitutions. The class B maltoporins are still able to form channels but display some specific features and altered specificity for maltose and maltodextrins. The substitution (Gly18----Val) alters trimer stability and impedes pore function (class C mutant). Thus, there is a good correlation between the specific transport properties of the mutated maltoporins in vivo and their behavior in vitro. These data, in combination with the asymmetric orientation of the protein within the bilayer and topological considerations, indicate that residues 245 and 163 do not belong to the selectivity filter. Mutations at these sites cause hindrance at the mouth of the pore on the outer domain of maltoporin.
Dargent
B
B
European Molecular Biology Laboratory, Heidelberg, F.R.G.
Charbit
A
A
Hofnung
M
M
Pattus
F
F
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D010582:Bacteriophage lambda / Q000378:metabolism; D001692:Biological Transport; D004926:Escherichia coli / Q000378:metabolism; D046911:Macromolecular Substances; D008320:Maltose / Q000378:metabolism; D009154:Mutation; D011134:Polysaccharides / Q000378:metabolism; D018272:Porins; D011991:Receptors, Virus / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D046911:Macromolecular Substances; D011134:Polysaccharides; D018272:Porins; D011991:Receptors, Virus; C016532:maltoporins; D008320:Maltose; C008315:maltodextrin