Location of three active site residues in the NH2-terminal sequence of the beta 2 subunit tryptophan synthase from Escherichia coli.
The three known active site residues of the tryptophan synthase beta 2 subunit from Escherichia coli are shown to fall within 25 residues of each other in the primary sequence of the NH2-terminal region of the beta 2 subunit. These residues are: lysine-86, which forms a Schiff's base with pyridoxal phosphate; histidine-81 or histidine-85, which removes the alpha proton of L-serine; and cysteine-61, which reacts with bromoacetylpyridoxamine phosphate, an affinity label for the beta 2 subunit. The sequence of the first 78 residues of a single cyanogen bromide fragment containing these active site residues has been determined by automatic Edman degradation and by sequence analysis of daughter peptides. This 79-residue cyanogen bromide fragment, which contains the 22-residue pyridoxyl peptide sequenced earlier by Fluri et al. (Fluri, R., Jackson, L. E., Lee, W. E., and Crawford, I. P. (1971) J. Biol. Chem. 246, 6620-6624), was placed at the NH2-terminal end of the beta chain beginning at residue 22. Thus, the primary sequence of residues 1 to 99 of the beta 2 subunit is reported.
D000595:Amino Acid Sequence; D001665:Binding Sites; D003488:Cyanogen Bromide; D004926:Escherichia coli / Q000201:enzymology*; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000032:analysis; D014357:Trypsin; D014367:Tryptophan Synthase / Q000378:metabolism
D046911:Macromolecular Substances; D010446:Peptide Fragments; D014357:Trypsin; D014367:Tryptophan Synthase; D003488:Cyanogen Bromide