Control of peptide deformylase activity by metal cations

Previous work indicated that peptide deformylase behaves as a metalloenzyme since the Escherichia coli enzyme was shown to copurify with a zinc ion. The present study establishes that nickel:enzyme complexes can also be isolated provided that nickel salts were added in the buffers throughout the purification. Similar results were obtained with the deformylases from Thermus thermophilus and Bacillus stearothermophilus. As a result of nickel binding, the catalytic efficiencies of peptide deformylases increased by two to three orders of magnitude with respect to those of the forms previously characterized. Using the model substrate N-formyl-Met-Ala-Ser, kcat/Km values of 5.4, 1.2 and 25 10(4)M-1s-1 could be obtained for the E. coli, T. thermophilus and B. stearothermophilus enzymes, respectively. This value satisfyingly accounts for the deformylation turnover required in the cell. In vitro characterization of the E. coli enzyme shows that zinc can readily substitute for the bound nickel with the catalytic efficiency decreasing to 80 M-1s-1 in turn. Conversely, the activity of the zinc-containing protein can be significantly improved by addition of nickel to the enzymatic assay.