Identification of the active-site lysine residues of two biosynthetic 3-dehydroquinases.
The lysine residues involved in Schiff-base formation at the active sites of both the 3-dehydroquinase component of the pentafunctional arom enzyme of Neurospora crassa and of the monofunctional 3-dehydroquinase of Escherichia coli were labelled by treatment with 3-dehydroquinate in the presence of NaB3H4. Radioactive peptides were isolated by h.p.l.c. following digestion with CNBr (and in one case after further digestion with trypsin). The sequence established for the N. crassa peptide was ALQHGDVVKLVVGAR, and that for the E. coli peptide was QSFDADIPKIA. An amended nucleotide sequence for the E. coli gene (aroD) that encode 3-dehydroquinase is also presented, along with a revised alignment of the deduced amino acid sequences for the biosynthetic enzymes.
Chaudhuri
S
S
Department of Biochemistry, University of Glasgow, Scotland, U.K.
Duncan
K
K
Graham
L D
LD
Coggins
J R
JR
D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D004926:Escherichia coli / Q000201:enzymology; D006836:Hydro-Lyases / Q000737:chemistry*; D008239:Lysine; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000201:enzymology; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D014316:Tritium
D010446:Peptide Fragments; D014316:Tritium; D006836:Hydro-Lyases; C020000:3-dehydroquinate dehydratase; D008239:Lysine; D003488:Cyanogen Bromide