Long-chain fatty acid transport in Escherichia coli. Cloning, mapping, and expression of the fadL gene

P. N. Black, S. F. Kianian, C. C. DiRusso, W. D. Nunn

Research output: Contribution to journalArticle

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Abstract

Transport of long-chain fatty acids across the inner membrane of Escherichia coli K-12 requires a functional fadL gene (Maloy, S.R., Ginsburgh, C.L., Simons, R.W., and Nunn, W.D. (1981) J. Biol. Chem. 256, 3735-3742). Mutants defective in the fadL gene lack a 33,000-dalton inner membrane protein as evaluated using two-dimensional pI/sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (Ginsburgh, C.L., Black, P.N., and Nunn, W.D. (1984) J. Biol. Chem. 259, 8437-8443). In an effort to determine whether the fadL gene is the structural gene for this 33,000-dalton protein, we have cloned, mapped, and analyzed the expression of the fadL gene. The fadL gene has been localized on a 2.8-kilobase EcoRV fragment of E. coli genomic DNA. Plasmids containing this gene (i) complement all fadL mutants, (ii) increase the long-chain fatty acid transport activity of fadL strains harboring them by 2- to 3-fold, and (iii) direct the synthesis of a membrane protein which has the same molecular weight and isoelectric point as that described by Ginsburgh et al. This is a heat-modifiable protein which has an apparent molecular weight of 43,000 daltons when solubilized at 100°C in the presence of SDS and 33,000 daltons when solubilized at 50°C in the presence of SDS.

Original languageEnglish (US)
Pages (from-to)1780-1789
Number of pages10
JournalJournal of Biological Chemistry
Volume260
Issue number3
StatePublished - Jan 1 1985

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Cloning
Escherichia coli
Organism Cloning
Fatty Acids
Genes
Gene Expression
Sodium Dodecyl Sulfate
Membrane Proteins
Molecular Weight
Molecular weight
Isoelectric Point
Electrophoresis
Polyacrylamide Gel Electrophoresis
Proteins
Plasmids
Hot Temperature
Membranes
DNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Long-chain fatty acid transport in Escherichia coli. Cloning, mapping, and expression of the fadL gene. / Black, P. N.; Kianian, S. F.; DiRusso, C. C.; Nunn, W. D.

In: Journal of Biological Chemistry, Vol. 260, No. 3, 01.01.1985, p. 1780-1789.

Research output: Contribution to journalArticle

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