Study of anthranilate synthase function by replacement of cysteine 84 using site-directed mutagenesis.
Cysteine 84 was replaced by glycine in Serratia marcescens anthranilate synthase Component II using site-directed mutagenesis of cloned trpG. This replacement abolished the glutamine-dependent anthranilate synthase activity but not the NH3-dependent activity of the enzyme. The mutation provides further evidence for the role of active site cysteine 84 in the glutamine amide transfer function of anthranilate synthase Component II. By the criteria of circular dichroism, proteolytic inactivation, and feedback inhibition the mutant and wild type enzymes were structurally similar. The NH3-dependent anthranilate synthase activity of the mutant enzyme supported tryptophan synthesis in media containing a high concentration of ammonium ion.
D000595:Amino Acid Sequence; D000641:Ammonia / Q000494:pharmacology; D000878:Anthranilate Synthase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D003545:Cysteine; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D005973:Glutamine / Q000494:pharmacology; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes; D010957:Plasmids; D011487:Protein Conformation; D012706:Serratia marcescens / Q000201:enzymology* / Q000235:genetics; D013329:Structure-Activity Relationship
D004269:DNA, Bacterial; D009692:Nucleic Acid Heteroduplexes; D005973:Glutamine; D000641:Ammonia; D000878:Anthranilate Synthase; D003545:Cysteine