title: Is the first enzyme of the shikimate pathway, 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (tyrosine sensitive), a copper metalloenzyme?
abstract: 3-Deoxy-D-arabino-heptulosonate-7-phosphate synthase (tyrosine sensitive) was purified from Escherichia coli carrying the plasmid pKB45. Enzyme of high specific catalytic activity (70 mu/mg) was obtained from cells grown only to the early log phase. The purified protein contained Cu(II) and showed an absorption band at 350 nm. Metal-free, catalytically inactive apoenzyme could be produced by dialysis against cyanide ion, and the holoenzyme could be reconstituted in terms of both catalytic activity and A350 by the binding of one Cu(II) ion per enzyme subunit. Zn(II) also reactivated the apoenzyme to about 50% of the level seen with Cu(II), although in this case no band appeared at 350 nm. In contrast to earlier reports that the enzyme contains substoichiometric levels of iron, insignificant amounts of iron were found in the isolated enzyme, and neither Fe(II) nor FE(III) regenerated either an absorption band at 350 nm or any catalytic activity from the apoenzyme. The evident preference of the enzyme as isolated for (Cu)II suggests that the synthase might naturally be a copper metalloenzyme.
authors.affiliation: Department of Chemistry, Harvard University, Cambridge, Massachusetts 02138.
authors.forename: T
authors.initials: T
authors.lastname: Baasov
authors.forename: J R
authors.initials: JR
authors.lastname: Knowles
mesh_terms: D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000302:isolation & purification / Q000378:metabolism*; D000446:Aldehyde-Lyases / Q000378:metabolism*; D001051:Apoenzymes / Q000302:isolation & purification / Q000378:metabolism; D003300:Copper / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D008667:Metalloproteins / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids; D013053:Spectrophotometry
chemical_list: D001051:Apoenzymes; D008667:Metalloproteins; D003300:Copper; D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D000446:Aldehyde-Lyases