Substitution of an arginyl residue for the active site lysyl residue (Lys258) of aspartate aminotransferase.
The active site lysyl residue (Lys258) of E. coli aspartate amino transferase was substituted for an arginyl residue by oligonucleotide-directed, site-specific mutagenesis. The mutant enzyme was obviously unable to form an aldimine bond with pyridoxal 5'-phosphate but firmly bound the coenzyme. The finding that the mutation did not lead to entire loss in the enzymic activity suggests that Lys258 may not be essential but auxiliary for enzymic catalysis. It is also conceived that the positive charge provided by Arg258 may contribute to the enzymic catalysis.
D000595:Amino Acid Sequence; D001120:Arginine; D001219:Aspartate Aminotransferases / Q000378:metabolism*; D001665:Binding Sites; D002942:Circular Dichroism; D004926:Escherichia coli / Q000201:enzymology; D008239:Lysine; D009154:Mutation; D011732:Pyridoxal Phosphate / Q000378:metabolism; D013053:Spectrophotometry; D013329:Structure-Activity Relationship
D011732:Pyridoxal Phosphate; D001120:Arginine; D001219:Aspartate Aminotransferases; D008239:Lysine