Site-specific alteration of cysteine 176 and cysteine 234 in the lactose carrier of Escherichia coli

R. J. Brooker, T. H. Wilson

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

In the present study, Cys-176 and Cys-234 in the lactose carrier have been modified to serine residues via site-specific mutagenesis. The resultant mutants have been characterized with regard to galactoside transport activity and sulfhydryl reagent sensitivity. The mutant proteins (in which Cys-176 or Cys-234 had been replaced with serine) are able to effectively transport galactosides, although the transport rates for lactose and methyl-β-D-galactopyranoside are slightly reduced compared to the normal lactose carrier. In addition, both mutants are less sensitive than the wild-type to high concentrations of two different sulfhydryl reagents, N-ethylmaleimide and p-hydroxymercuribenzoate. Overall, the data are consistent with the idea that Cys-176 and Cys-234 are close to the substrate recognition site. However, neither residue appears to be essential for galactoside transport by providing an ionizable group near the active site or by forming a disulfide bond.

Original languageEnglish (US)
Pages (from-to)11765-11769
Number of pages5
JournalJournal of Biological Chemistry
Volume261
Issue number25
StatePublished - Dec 1 1986

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Galactosides
Lactose
Escherichia coli
Cysteine
Sulfhydryl Reagents
Serine
Mutagenesis
Ethylmaleimide
Mutant Proteins
Site-Directed Mutagenesis
Galactose
Disulfides
Catalytic Domain
Substrates

Cite this

Site-specific alteration of cysteine 176 and cysteine 234 in the lactose carrier of Escherichia coli. / Brooker, R. J.; Wilson, T. H.

In: Journal of Biological Chemistry, Vol. 261, No. 25, 01.12.1986, p. 11765-11769.

Research output: Contribution to journalArticle

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