Isolation of glutamic acid methyl ester from an Escherichia coli membrane protein involved in chemotaxis

S. J. Kleene, Myron Lee Toews, J. Adler

Research output: Contribution to journalArticle

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Abstract

Glutamic acid 5-methyl ester was isolated from an E. coli protein involved in chemotaxis. The bacteria were first incubated with [methyl-3H] methionine under conditions which are known to result in methylation of the protein. The protein, isolated by gel electrophoresis, was then digested by successive treatment with three proteolytic enzymes. One of the products was [methyl-3H] glutamic acid 5-methyl ester, identified by comparison with an authentic sample in the following studies: chromatography on an automatic amino acid analyzer, chromatography on paper in two solvent systems, chromatography on paper of the N-acetyl derivatives, and stability of the ester bond to various pH conditions. No aspartic acid 4-methyl ester was found in the enzymatic digest. Treatment of the methylated protein with alkali released the radioactivity as [3H] methanol, which was identified by gas chromatography and by preparation of the 3,5-dinitrobenzoate.

Original languageEnglish (US)
Pages (from-to)3214-3218
Number of pages5
JournalJournal of Biological Chemistry
Volume252
Issue number10
StatePublished - Jan 1 1977

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Escherichia coli Proteins
Chemotaxis
Chromatography
Escherichia coli
Glutamic Acid
Paper Chromatography
Membrane Proteins
Esters
Proteins
Methylation
Radioactivity
Alkalies
Electrophoresis
Gas chromatography
Methionine
Gas Chromatography
Methanol
Bacteria
Peptide Hydrolases
Gels

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Isolation of glutamic acid methyl ester from an Escherichia coli membrane protein involved in chemotaxis. / Kleene, S. J.; Toews, Myron Lee; Adler, J.

In: Journal of Biological Chemistry, Vol. 252, No. 10, 01.01.1977, p. 3214-3218.

Research output: Contribution to journalArticle

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