Escherichia coli gamma-glutamyltranspeptidase mutants deficient in processing to subunits.
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.
Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
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Hashimoto
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Suzuki
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Nohara
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Kumagai
D000409:Alanine; D000595:Amino Acid Sequence; D001120:Arginine; D001483:Base Sequence; D015153:Blotting, Western; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005838:Genotype; D005998:Glycine; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D005723:gamma-Glutamyltransferase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D001120:Arginine; D005723:gamma-Glutamyltransferase; D000409:Alanine; D005998:Glycine