A five-coordinate metal center in Co(II)-substituted VanX

被引:30
作者
Breece, RM
Costello, A
Bennett, B
Sigdel, TK
Matthews, ML
Tierney, DL
Crowder, MW
机构
[1] Miami Univ, Dept Chem & Biochem, Oxford, OH 45056 USA
[2] Univ New Mexico, Dept Chem, Albuquerque, NM 87131 USA
[3] Med Coll Wisconsin, Natl Biomed EPR Ctr, Dept Biophys, Milwaukee, WI 53226 USA
关键词
D O I
10.1074/jbc.M412582200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 similar to 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 = vertical bar +/- 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 similar to 2.0 angstrom. 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.
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页码:11074 / 11081
页数:8
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