Mechanism of sugar transport through the sugar-specific LamB channel ofEscherichia coli outer membrane

SummaryLipid bilayer experiments were performed with the sugar-specific LamB (maltoporin) channel ofEscherichia coli outer membrane. Single-channel analysis of the conductance steps caused by LamB showed that there was a linear relationship between the salt concentration in the aqueous phase and the channel conductance, indicating only small or no binding between the ions and the channel interior. The total or the partial blockage of the ion movement through the LamB channel was not dependent on the ion concentration in the aqueous phase. Both results allowed the investigation of the sugar binding in more detail, and the stability constants of the binding of a large variety of sugars to the binding site inside the channel were calculated from titration experiments of the membrane conductance with the sugars. The channel was highly cation selective, both in the presence and absence of sugars, which may be explained by the existence of carbonyl groups inside the channel. These carbonyl groups may also be involved in the sugar binding via hydrogen bonds. The kinetics of the sugar transport through the LamB channel were estimated relative to maltose by assuming a simple one-site, two-barrier model from the relative rates of permeation taken from M. Luckey and H. Nikaido (Proc. Natl. Acad. Sci. USA77:165–171 (1980a)) and the stability constants for the sugar binding given in this study.

[1]  R. Benz,et al.  Ionic selectivity of pores formed by the matrix protein (porin) of Escherichia coli. , 1979, Biochimica et biophysica acta.

[2]  H. Nikaido,et al.  Specificity of diffusion channels produced by X phage receptor protein of Escherichia coli (liposomes/reconstitution/outer membrane/facilitated diffusion/maltose) , 1980 .

[3]  E. T. Palva,et al.  Major outer membrane protein in Salmonella typhimurium induced by maltose , 1978, Journal of bacteriology.

[4]  R. Benz,et al.  Mechanism of ion transport through the anion-selective channel of the Pseudomonas aeruginosa outer membrane , 1987, The Journal of general physiology.

[5]  T. Ferenci,et al.  Directed evolution of the lambda receptor of Escherichia coli through affinity chromatographic selection. , 1982, Journal of molecular biology.

[6]  M. Hofnung,et al.  Maltose transport in Escherichia coli K-12: involvement of the bacteriophage lambda receptor , 1975, Journal of bacteriology.

[7]  H. Nikaido,et al.  Outer membrane of gram-negative bacteria. XVIII. Electron microscopic studies on porin insertion sites and growth of cell surface of Salmonella typhimurium , 1978, Journal of bacteriology.

[8]  R. Crane Intestinal absorption of sugars. , 1960, Physiological reviews.

[9]  G. Kimmich Coupling between Na+ and sugar transport in small intestine. , 1973, Biochimica et biophysica acta.

[10]  H. Nikaido,et al.  Outer membrane of gram-negative bacteria. XVII. Secificity of transport process catalyzed by the lambda-receptor protein in Escherichia coli. , 1977, Biochemical and biophysical research communications.

[11]  H. Nikaido,et al.  Porin channels in Escherichia coli: studies with liposomes reconstituted from purified proteins , 1983, Journal of bacteriology.

[12]  R. Benz,et al.  Ion selectivity of gram-negative bacterial porins , 1985, Journal of bacteriology.

[13]  R. Benz,et al.  Outer membrane protein P of Pseudomonas aeruginosa: regulation by phosphate deficiency and formation of small anion-specific channels in lipid bilayer membranes , 1982, Journal of bacteriology.

[14]  R. Latorre,et al.  Conduction and selectivity in potassium channels , 2005, The Journal of Membrane Biology.

[15]  R. Benz,et al.  Properties of the large ion-permeable pores formed from protein F of Pseudomonas aeruginosa in lipid bilayer membranes. , 1981, Biochimica et biophysica acta.

[16]  M. Hofnung,et al.  High-sensitivity detection of newly induced LamB protein on the Escherichia coli cell surface , 1984, Journal of bacteriology.

[17]  P. Läuger Ion transport through pores: a rate-theory analysis. , 1973, Biochimica et biophysica acta.

[18]  R. Benz,et al.  Pore formation by LamB of Escherichia coli in lipid bilayer membranes , 1986, Journal of bacteriology.

[19]  R. Benz,et al.  Porin from bacterial and mitochondrial outer membranes. , 1985, CRC critical reviews in biochemistry.

[20]  R. Benz,et al.  Pore-forming activity of the Tsx protein from the outer membrane of Escherichia coli. Demonstration of a nucleoside-specific binding site. , 1988, The Journal of biological chemistry.

[21]  A. Irimajiri,et al.  Electrical properties and active solute transport in rat small intestine , 1977, The Journal of Membrane Biology.

[22]  T. Ferenci,et al.  The role of the maltodextrin-binding site in determining the transport properties of the LamB protein. , 1986, Journal of Biological Chemistry.

[23]  H. Nikaido,et al.  Diffusion of solutes through channels produced by phage lambda receptor protein of Escherichia coli: inhibition by higher oligosaccharides of maltose series. , 1980, Biochemical and biophysical research communications.

[24]  H. Nikaido,et al.  Molecular basis of bacterial outer membrane permeability. , 1985, Microbiological reviews.

[25]  R. Benz,et al.  Formation of large, ion-permeable membrane channels by the matrix protein (porin) of Escherichia coli. , 1978, Biochimica et biophysica acta.

[26]  T. Nakae,et al.  Identification of the outer membrane protein of E. coli that produces transmembrane channels in reconstituted vesicle membranes. , 1976, Biochemical and biophysical research communications.

[27]  A. Finkelstein,et al.  The gramicidin a channel: A review of its permeability characteristics with special reference to the single-file aspect of transport , 1981, The Journal of Membrane Biology.

[28]  T. Ferenci,et al.  Sequence determinants in the lamB gene of Escherichia coli influencing the binding and pore selectivity of maltoporin. , 1987, Gene.

[29]  R. Benz,et al.  Properties of chemically modified porin from Escherichia coli in lipid bilayer membranes. , 1984, Biochimica et biophysica acta.

[30]  T. Ferenci,et al.  Lambda Receptor in the Outer Membrane of Escherichia coli as a Binding Protein for Maltodextrins and Starch Polysaccharides , 1980, Journal of bacteriology.

[31]  R. Hancock Role of porins in outer membrane permeability , 1987, Journal of bacteriology.

[32]  W. R. Lieb,et al.  Testing and characterizing the simple pore. , 1974, Biochimica et biophysica acta.