Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 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.

Full text

PDF
129

Selected References

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

  1. ALBERT A. Quantitative studies of the avidity of naturally occurring substances for trace metals; amino-acids having only two ionizing groups. Biochem J. 1950 Nov-Dec;47(5):531–538. doi: 10.1042/bj0470531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BRUNO G. A., CHRISTIAN J. E. Note on suitable solvent systems usable in the liquid scintillation counting of animal tissue. J Am Pharm Assoc Am Pharm Assoc. 1960 Aug;49:560–561. doi: 10.1002/jps.3030490820. [DOI] [PubMed] [Google Scholar]
  3. Dalgleish D. G., Peacocke A. R. Circular-dichroism studies on two -lactamases from Bacillus cereus. Biochem J. 1971 Nov;125(1):155–158. doi: 10.1042/bj1250155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Davies R. B., Abraham E. P. Separation, purification and properties of beta-lactamase I and beta-lactamase II from Bacillus cereus 569/H/9. Biochem J. 1974 Oct;143(1):115–127. doi: 10.1042/bj1430115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Drum D. E., Vallee B. L. Optical properties of catalytically active cobalt and cadmium liver alcohol dehydrogenases. Biochem Biophys Res Commun. 1970 Oct 9;41(1):33–39. doi: 10.1016/0006-291x(70)90464-x. [DOI] [PubMed] [Google Scholar]
  6. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  7. Good N. E., Winget G. D., Winter W., Connolly T. N., Izawa S., Singh R. M. Hydrogen ion buffers for biological research. Biochemistry. 1966 Feb;5(2):467–477. doi: 10.1021/bi00866a011. [DOI] [PubMed] [Google Scholar]
  8. Kuwabara S., Adams E. P., Abraham E. P. The composition of beta-lactamase I and beta-lactamase II from Bacillus cereus 569-H. Biochem J. 1970 Jul;118(3):475–480. doi: 10.1042/bj1180475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. MALMSTROM B. G. Determination of zinc in biological materials. Methods Biochem Anal. 1956;3:327–352. [PubMed] [Google Scholar]
  10. MYER Y. P., SCHELLMAN J. A. The interaction of ribonuclease with purine and pyrimidine phosphates. I. Binding of adenosine 5'-monophosphate to ribonuclease. Biochim Biophys Acta. 1962 Mar 5;55:361–373. doi: 10.1016/0006-3002(62)90791-6. [DOI] [PubMed] [Google Scholar]
  11. NEWTON G. G., ABRAHAM E. P. Isolation of cephalosporin C, a penicillin-like antibiotic containing D-alpha-aminoadipic acid. Biochem J. 1956 Apr;62(4):651–658. doi: 10.1042/bj0620651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sabath L. D., Finland M. Thiol-group binding of zinc to a beta-lactamase of Bacillus cereus: differential effects on enzyme activity with penicillin and cephalosporins as substrates. J Bacteriol. 1968 May;95(5):1513–1519. doi: 10.1128/jb.95.5.1513-1519.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Vallee B. L., Williams R. J. Enzyme action: views derived from metalloenzyme studies. Chem Br. 1968 Sep;4(9):397–402. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES