Active Transport by the CscB Permease in Escherichia coli K-12

The cscB gene encoding a putative sucrose permease from Escherichia coli EC3132 [Bockmann, J., Heuel, H. & Lengeler, J. W. (1992) Mol. Gen. Genet. 235, 22-32] was placed under control of the lacZ promoter/operator and overexpressed. Cells of E. coli K-12 expressing cscB gene product from a high...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1995-03, Vol.208 (3), p.1116-1123
Main Authors: Sahintoth, M., Frillingos, S., Lengeler, J.W., Kaback, H.R.
Format: Article
Language:English
Subjects:
Base Sequence
beta-Galactosidase - genetics
Biological Transport, Active
Cell Membrane - enzymology
Cloning, Molecular
DNA, Bacterial - chemistry
DNA, Bacterial - metabolism
Epitopes - analysis
Escherichia coli - enzymology
Escherichia coli - genetics
Escherichia coli Proteins
Gene Expression
Genes, Bacterial
Kinetics
Membrane Transport Proteins - biosynthesis
Membrane Transport Proteins - metabolism
Molecular Sequence Data
Plasmids
Promoter Regions, Genetic
Protein Biosynthesis
Sucrose - metabolism
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Summary:The cscB gene encoding a putative sucrose permease from Escherichia coli EC3132 [Bockmann, J., Heuel, H. & Lengeler, J. W. (1992) Mol. Gen. Genet. 235, 22-32] was placed under control of the lacZ promoter/operator and overexpressed. Cells of E. coli K-12 expressing cscB gene product from a high copy number plasmid accumulate sucrose against a concentration gradient with an apparent Km of about 1 mM and a Vmax of approximately 23 nmol/min/mg protein. In contrast, transport of lactose or melibiose is not observed. Epitope-tagging and immunological analysis reveal that concomitant with increased transport activity, a protein with an apparent Mr of 33-34 kDa appears in the cytoplasmic membrane. The results provide direct support for the contention that the cscB gene encodes a sucrose permease and provide the basis for further molecular analysis.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1995.1449