Medical College of Wisconsin
CTSICores SearchResearch InformaticsREDCap

A five-coordinate metal center in Co(II)-substituted VanX. J Biol Chem 2005 Mar 25;280(12):11074-81

Date

01/20/2005

Pubmed ID

15657055

DOI

10.1074/jbc.M412582200

Scopus ID

2-s2.0-15744379019 (requires institutional sign-in at Scopus site)   34 Citations

Abstract

In an effort to structurally probe the metal binding site in VanX, electronic absorption, EPR, and extended x-ray absorption fine structure (EXAFS) spectroscopic studies were conducted on Co(II)-substituted VanX. Electronic spectroscopy revealed the presence of Co(II) ligand field transitions that had molar absorptivities of approximately 100 m(-1) cm(-1), which suggests that Co(II) is five-coordinate in Co(II)-substituted VanX. Low temperature EPR spectra of Co(II)-substituted VanX were simulated using spin Hamiltonian parameters of M(S) = |+/-1/2), E/D = 0.14, g(real(x,y)) = 2.37, and g(real(z)) = 2.03. These parameters lead to the prediction that Co(II) in the enzyme is five-coordinate and that there may be at least one solvent-derived ligand. Single scattering fits of EXAFS data indicate that the metal ions in both native Zn(II)-containing and Co(II)-substituted VanX have the same coordination number and that the metal ions are coordinated by 5 nitrogen/oxygen ligands at approximately 2.0 angstroms. These data demonstrate that Co(II) (and Zn(II) from EXAFS studies) is five-coordinate in VanX in contrast to previous crystallographic studies (Bussiere, D. E., Pratt, S. D., Katz, L., Severin, J. M., Holzman, T., and Park, C. H. (1998) Mol. Cell 2, 75-84). These spectroscopic studies also demonstrate that the metal ion in Co(II)-substituted VanX when complexed with a phosphinate analog of substrate D-Ala-D-Ala is also five-coordinate.

Author List

Breece RM, Costello A, Bennett B, Sigdel TK, Matthews ML, Tierney DL, Crowder MW

Author

Brian Bennett D.Phil. Professor and Chair in the Physics department at Marquette University




MESH terms used to index this publication - Major topics in bold

Bacterial Proteins
Binding Sites
Cobalt
Crystallography
Electron Spin Resonance Spectroscopy
Serine-Type D-Ala-D-Ala Carboxypeptidase
Zinc