Abstract
Arginyl residues 513 and 571 of Escherichia coli K-12 gamma-glutamyl-transpeptidase (EC 2.3.2.2) were substituted with alanyl and glycyl residues, respectively, by oligonucleotide-directed in vitro mutagenesis. Both mutants were devoid of the enzymatic activity. On Western blot analysis, we found that both mutants accumulated a gamma-glutamyltranspeptidase precursor which was not processed into large and small subunits in the periplasmic space of Escherichia coli.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alanine
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Amino Acid Sequence
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Arginine
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Base Sequence
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Blotting, Western
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Genotype
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Glycine
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Kinetics
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Macromolecular Substances
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Molecular Sequence Data
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Mutagenesis, Site-Directed*
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Oligodeoxyribonucleotides
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Restriction Mapping
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gamma-Glutamyltransferase / genetics*
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gamma-Glutamyltransferase / isolation & purification
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gamma-Glutamyltransferase / metabolism*
Substances
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Macromolecular Substances
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Oligodeoxyribonucleotides
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Recombinant Proteins
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Arginine
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gamma-Glutamyltransferase
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Alanine
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Glycine