title: Isolation and crystallization of functionally competent Escherichia coli peptide deformylase forms containing either iron or nickel in the active site.
abstract: Three metallo forms of peptide deformylase (PDF, EC 3.5.1.31) of Escherichia coli were prepared and crystallized (space group C2, diffraction limit 1.9 A) for initiating the X-ray structure determination of the metal center in correlation with the catalytic functionality of this enzyme. The native Fe2+ containing enzyme species was directly isolated from overproducing bacteria by using catalase as a buffer additive, which stabilizes the catalytic activity against oxidative destruction. The Ni2+ containing form, which is oxygen-insensitive, was obtained by metal exchange with free Ni2+ and found to be catalytically equally effective (kcat/KM = 10(5) M-1 s-1 for N-formyl-Met-Ala). The Zn2+ form, prepared from the apoenzyme or by displacement of bound Ni2+ by free Zn2+, proved virtually inactive.
authors.affiliation: Biochemie-Zentrum Heidelberg, Ruprecht-Karls Universität, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Groche
authors.forename: A
authors.initials: A
authors.lastname: Becker
authors.forename: I
authors.initials: I
authors.lastname: Schlichting
authors.forename: W
authors.initials: W
authors.lastname: Kabsch
authors.forename: S
authors.initials: S
authors.lastname: Schultz
authors.forename: A F
authors.initials: AF
authors.lastname: Wagner
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001665:Binding Sites; D002374:Catalase; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D007501:Iron / Q000737:chemistry; D007700:Kinetics; D009532:Nickel / Q000737:chemistry; D010084:Oxidation-Reduction; D015032:Zinc / Q000737:chemistry
chemical_list: D009532:Nickel; D007501:Iron; D002374:Catalase; D000581:Amidohydrolases; C020150:formylmethionine deformylase; D015032:Zinc