DOI: 10.1111/j.1432-1033.1988.tb14333.x. Characterization of the nucleoside‐binding site inside the Tsx channel of<i>Escherichia coli</i>outer membrane Reconstitution experiments with lipid bilayer membranes

Characterization of the nucleoside‐binding site inside the Tsx channel ofEscherichia coliouter membrane Reconstitution experiments with lipid bilayer membranes

10.1111/j.1432-1033.1988.tb14333.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

Reconstitution of purified Tsx protein fromEscherichia coliinto lipid bilayer membranes showed that Tsx formed small ion‐permeable channels with a single‐channel conductance of 10 pS in 1 M KCI. The dependence of conductance versus salt concentration was linear, suggesting that Tsx has no binding site for ions. Conductance was inhibited by the addition of 20 mM adenosine. Titration of the Tsx‐mediated membrane conductance with different solutes including free bases, nucleosides, and deoxynucleosides suggested that the channel contains a binding site for nucleosides but not for sugars or amino acids, and binding increased in the following order: free base, nucleoside, and deoxynucleoside. Among the five nucleosides the stability constant for the binding increased in the order of cytidine, guanosine, uridine, adenosine, and thymidine. Control experiments revealed that the binding of the nucleosides is independent of ion concentration in the aqueous phase, i.e. there was no competition between nucleosides and ions for the binding site inside the channel. The binding of the solutes to the channel interior can be explained by a one‐site two‐barrier model for the Tsx channel. The advantage of a binding site inside a specific porin for the permeation of solutes is discussed with respect to the properties of a general diffusion pore.

Bibliography

BENZ, R., SCHMID, A., MAIER, C., & BREMER, E. (1988). Characterization of the nucleosideâbinding site inside the Tsx channel ofEscherichia coliouter membrane Reconstitution experiments with lipid bilayer membranes. European Journal of Biochemistry, 176(3), 699â705. Portico.

Authors 4

Roland BENZ (first)
Angela SCHMID (additional)
Christl MAIER (additional)
Erhard BREMER (additional)

References 44 Referenced 47

{'key': 'e_1_2_1_2_2', 'first-page': '1011', 'volume': '143', 'author': 'Fredericq F.', 'year': '1949', 'journal-title': 'C. R. Soc. Biol.'} / C. R. Soc. Biol. by Fredericq F. (1949)
10.1084/jem.133.3.534
10.1016/0006-291X(76)90810-X
{'key': 'e_1_2_1_5_2', 'first-page': '203', 'volume-title': 'Metabolism of nucleotides, nucleosides and nucleobases in microorganisms', 'author': 'Hammer‐Jespersen K.', 'year': '1983'} / Metabolism of nucleotides, nucleosides and nucleobases in microorganisms by Hammer‐Jespersen K. (1983)
10.1007/BF00427732
10.1128/jb.170.1.108-116.1988
10.1016/0014-5793(76)80737-5
10.1128/jb.126.2.814-822.1976 / J. Bacteriol. by McKenow M. (1976)
10.1016/S0021-9258(18)50647-5 / J. Biol. Chem. by Munch‐Petersen A. (1979)
10.1128/jb.131.2.631-637.1977
10.1016/0014-5793(78)81264-2
10.1007/BF00401750
10.3109/10409238509082542
10.1128/mr.49.1.1-32.1985
10.1128/jb.169.3.929-933.1987 / J. Bacteriol. by Hancook R. E. W. (1987)
10.1007/978-3-642-71399-6_1
{'key': 'e_1_2_1_18_2', 'first-page': '259', 'volume-title': 'Metabolism of nucleotides, nucleosides and nucleobases in microorganisms', 'author': 'Munch‐Petersen A.', 'year': '1983'} / Metabolism of nucleotides, nucleosides and nucleobases in microorganisms by Munch‐Petersen A. (1983)
{'key': 'e_1_2_1_19_2', 'first-page': '445', 'volume-title': 'Escherichia coli and Salmonella typhimurium', 'author': 'Neuhard G.', 'year': '1987'} / Escherichia coli and Salmonella typhimurium by Neuhard G. (1987)
10.1016/S0021-9258(18)69233-6
{'key': 'e_1_2_1_21_2', 'first-page': '1482', 'volume-title': 'Escherichia coli and Salmonella typhimurium', 'author': 'Schwartz M.', 'year': '1987'} / Escherichia coli and Salmonella typhimurium by Schwartz M. (1987)
10.1128/jb.124.1.112-118.1975
10.1111/j.1432-1033.1976.tb10383.x
10.1073/pnas.77.1.167
10.1016/S0006-291X(80)80261-0
10.1016/S0021-9258(17)36137-9 / J. Biol. Chem. by Nakae T.
10.1128/jb.142.2.521-526.1980
10.1128/jb.165.3.978-986.1986
10.1016/S0021-9258(19)57394-X
{'key': 'e_1_2_1_30_2', 'first-page': '12', 'volume': '100', 'author': 'Benz R.', 'year': '1986', 'journal-title': 'J. Memb. Biol.'} / J. Memb. Biol. by Benz R. (1986)
10.1016/0006-291X(77)90534-4
10.1128/jb.147.3.1113-1116.1981 / J. Bacteriol. by Heuzenroeder M. W. (1981)
10.1128/JB.119.3.726-735.1974
{'key': 'e_1_2_1_34_2', 'volume-title': 'Experiments in molecular genetics', 'author': 'Miller J. H.', 'year': '1972'} / Experiments in molecular genetics by Miller J. H. (1972)
10.1016/0014-5793(75)80272-9
10.1038/227680a0
10.1016/0003-2697(81)90179-2
10.1085/jgp.89.2.275
10.1016/0005-2736(89)90002-3 / Biochim. Biophys. Acta by Benz R (1979)
10.1016/0005-2736(89)90002-3
10.1007/BF01869348
{'key': 'e_1_2_1_42_2', 'first-page': '317', 'volume-title': 'Achievements and perspectives of mitochondrial research', 'author': 'Benz R.', 'year': '1985'} / Achievements and perspectives of mitochondrial research by Benz R. (1985)
10.1128/jb.162.2.722-727.1985
10.1016/0014-5793(87)80891-8
10.1016/0005-2736(86)90569-9

Dates

Type	When
Created	20 years, 6 months ago (March 3, 2005, 9:21 p.m.)
Deposited	1 year, 7 months ago (Jan. 23, 2024, 6:22 p.m.)
Indexed	1 year, 2 months ago (July 1, 2024, 6:31 p.m.)
Issued	36 years, 11 months ago (Oct. 1, 1988)
Published	36 years, 11 months ago (Oct. 1, 1988)
Published Online	20 years, 6 months ago (March 3, 2005)
Published Print	36 years, 11 months ago (Oct. 1, 1988)

Funders 0

None

BibTeX

@article{BENZ_1988, title={Characterization of the nucleoside‐binding site inside the Tsx channel ofEscherichia coliouter membrane Reconstitution experiments with lipid bilayer membranes}, volume={176}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1988.tb14333.x}, DOI={10.1111/j.1432-1033.1988.tb14333.x}, number={3}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={BENZ, Roland and SCHMID, Angela and MAIER, Christl and BREMER, Erhard}, year={1988}, month=oct, pages={699–705} }

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