Preference of Cd(II) and Zn(II) for the two metal sites in Bacillus cereus beta-lactamase II: A perturbed angular correlation of gamma-rays spectroscopic study.
Paul-Soto, R.; Zeppezauer, M.; Adolph, H. W.et al.
[en] Cd-substituted forms of the Bacillus cereus metallo-beta-lactamases (BCII) were studied by perturbed angular correlation of gamma-rays (PAC) spectroscopy. At very low [Cd]:[apo-beta-lactamase] ratios, two nuclear quadrupole interactions (NQI) were detected. For [Cd]:[apo-beta-lactamase] ratios between 0.8 and 3.0, two new NQIs appear, and the spectra show that up to 2 cadmium ions can be bound per molecule of apoenzyme. These results show the existence of two interacting Cd-binding sites in BCII. The relative populations of the two NQIs found at low [Cd]:[apo-beta-lactamase] ratios yielded a 1:3 ratio for the microscopic dissociation constants of the two different metal sites (when only one cadmium ion is bound). X-ray diffraction data at pH 7.5 demonstrate that also for Zn(II) two binding sites exist, which may be bridged by a solvent molecule. The measured NQIs could be assigned to the site with three histidines as metal ligands (three-His site) and to the site with histidine, cysteine, and aspartic acid as metal ligands (Cys site), respectively, by PAC measurements on the Cys168Ala mutant enzyme. This assignment shows that cadmium ions preferentially bind to the Cys site. This is in contrast to the preference of Zn(II) in the hybrid Zn(II)Cd(II) enzyme, where an analysis of the corresponding PAC spectrum showed that Cd(II) occupied the Cys site, whereby Zn(II) occupied the site with three histidines. The difference between Zn(II) and Cd(II) in affinity for the two sites is combined with the kinetics of hydrolysis of nitrocefin for different metal ion substitutions (Zn(2)E, ZnE, Cd(2)E, CdE, and ZnCdE) to study the function of the two metal ion binding sites.
Frère, Jean-Marie ; Université de Liège > Département des sciences de la vie > Centre d'ingénierie des protéines
Carfi, A.
Dideberg, O.
Wouters, J.
Hemmingsen, L.
Bauer, R.
Language :
English
Title :
Preference of Cd(II) and Zn(II) for the two metal sites in Bacillus cereus beta-lactamase II: A perturbed angular correlation of gamma-rays spectroscopic study.
Publication date :
1999
Journal title :
Biochemistry
ISSN :
0006-2960
eISSN :
1520-4995
Publisher :
American Chemical Society, Washington, United States - District of Columbia
Carfi, A., Pares, S., Duée, E., Galleni, M., Duez, C., Frère, J.-M., and Dideberg, O. (1995) EMBO J. 14, 4914-4921.
Baldwin, G. S., Galdes, A., Hill, H. A. O., Smith, B. E., Waley, S. G., and Abraham, E. P. (1978) Biochem. J. 175, 441-447.
Baldwin, G. S., Waley, S. G., and Abraham, E. P. (1979) Biochem. J. 179, 459-463.
Baldwin, G. S., Galdes, A., Hill, H. A. O., Waley, S. G., and Abraham, E. P. (1980) J. Inorg. Biochem. 13, 189-204.
Carfi, A., Duée, E., Galleni, M., Frère, J.-M., and Dideberg, O. (1998) Acta Crystallogr. D54, 313-323.
Carfi, A., Duée, E., Paul-Soto, R., Galleni, M., Frère, J.-M., and Dideberg, O. (1998) Acta Crystallogr. D54, 45-57.
Concha, N. O., Rasmussen, B. A., Bush, K., and Herzberg, O. (1996) Structure 4, 823-836.
Concha, N. O., Rasmussen, B. A., Bush, K., and Herzberg, O. (1997) Protein Sci. 6, 2671-2676.
Carfi, A., Paul-Soto, R., Martin, T., Pétillot, Y., Frère, J.-M.; Dideberg, O. (1997) Acta Crystallogr. D53, 485-487.
Paul-Soto, R., Bauer, R., Frère, J. M., Galleni, M., Meyer-Klaucke, W., Nolting, H., Rossolini, G. M., de Seny, D., Hernandez-Valladares, M., Zeppezauer, M., and Adolph, H. W. (1999) J. Biol. Chem. 274, 13242-13249.
Adolph, H. W., Kiefer, M., and Cedergren-Zeppezauer, E. (1997) Biochemistry 36, 8743-8754.
Hemmingsen, L., Bauer, R., Bjerrum, M. J., Adolph, H. W., Zeppezauer, M., and Cedergren-Zeppezauer, E. (1996) Eur. J. Biochem. 241, 546-551.
Bauer, R., Danielsen, E., Hemmingsen L., Soørensen, M. V., Ulstrup, J., Friis, E. P., Auld, D. S., and Bjerrum, M. J. (1997) Biochemistry 36, 11514-11524.
Bauer, R. (1985) Q. Rev. Biophys. 18, 1-64.
Danielsen, E., Bauer, R., and Schneider, D. (1991) Eur. Biophys. J. 20, 193-201.
Paul-Soto, R., Hernandez-Valladares, M., Galleni, M., Bauer, R., Zeppezauer, M., Frère, J.-M., and Adolph, H. W. (1998) FEBS Lett. 438, 137-140.
Bauer, R., Limkilde, P., and Johansen, J. T. (1976) Biochemistry 15, 334-342.
Bauer, R., Jensen, S. J., and Schmidt-Nielsen, B., (1988) Hyperfine Interact. 39, 203-234.
Vallee, B. L., and Auld, D. S. (1993) Biochemistry 32, 6493-6500.
Bounaga, S., Laws, A. P., Galleni, M., and Page, M. I. (1998) Biochem. J. 331, 703-711.