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. 1974 Oct;143(1):129–135. doi: 10.1042/bj1430129

Metal cofactor requirement of β-lactamase II

Richard B Davies 1, E P Abraham 1
PMCID: PMC1168360  PMID: 4219279

Abstract

1. The apoenzyme obtained on removal of Zn2+ from β-lactamase II from Bacillus cereus 569/H/9 showed less than 0.001% of the activity of the Zn2+-containing enzyme. 2. Removal of Zn2+ led to a conformational change in the enzyme and partial unmasking of a thiol group. 3. Replacement of Zn2+ by Co2+, Cd2+, Mn2+ or Hg2+ gave enzymes with significant, but lower, β-lactamase activity. No activity was detected in the presence of Cu2+, Ni2+, Mg2+ or Ca2+. 4. Equilibrium dialysis indicated that the enzyme had at least two Zn2+ binding sites. With benzylpenicillin as substrate the variation in activity with concentration of Zn2+ indicated that activity paralleled binding of Zn2+ to the site of highest affinity. 5. Replacement of Zn2+ by Co2+ and Cd2+ gave enzymes with absorption bands at 340 and 245nm respectively, and raised the question of whether the thiol group in the enzyme is a metal-ion ligand. 6. Reduction of the product obtained by reaction of denatured β-lactamase II with Ellman's reagent [5,5′-dithiobis-(2-nitrobenzoic acid)] gave a protein which could refold to produce β-lactamase II activity in high yield.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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