PMID:16430228
| Citation |
Periyannan, GR, Costello, AL, Tierney, DL, Yang, KW, Bennett, B and Crowder, MW (2006) Sequential binding of cobalt(II) to metallo-beta-lactamase CcrA. Biochemistry 45:1313-20 |
|---|---|
| Abstract |
In an effort to probe Co(II) binding to metallo-beta-lactamase CcrA, EPR, EXAFS, and (1)H NMR studies were conducted on CcrA containing 1 equiv (1-Co(II)-CcrA) and 2 equiv (Co(II)Co(II)-CcrA) of Co(II). The EPR spectra of 1-Co(II)-CcrA and Co(II)Co(II)-CcrA are distinct and indicate 5/6-coordinate Co(II) ions. The EPR spectra also reveal the absence of significant spin-exchange coupling between the Co(II) ions in Co(II)Co(II)-CcrA. EXAFS spectra of 1-Co(II)-CcrA suggest 5/6-coordinate Co(II) with two or more histidine ligands. EXAFS spectra of Co(II)Co(II)-CcrA also indicate 5/6 ligands at a similar average distance to 1-Co(II)-CcrA, including an average of about two histidines per Co(II). (1)H NMR spectra for 1-Co(II)-CcrA revealed seven paramagnetically shifted resonances, three of which were solvent-exchangeable, while the NMR spectra for Co(II)Co(II)-CcrA showed at least 16 shifted resonances, including an additional solvent-exchangeable resonance and a resonance at 208 ppm. The data indicate sequential binding of Co(II) to CcrA and that the first Co(II) binds to the consensus Zn(1) site in the enzyme. |
| Links |
PubMed Online version:10.1021/bi051105n |
| Keywords |
Bacterial Proteins/chemistry; Bacterial Proteins/metabolism; Binding Sites; Cobalt/chemistry; Electron Spin Resonance Spectroscopy/methods; Escherichia coli/enzymology; Kinetics; Magnetic Resonance Spectroscopy/methods; Metals/chemistry; Metals/metabolism; Models, Molecular; Protein Binding; Spectrophotometry, Ultraviolet/methods; Spectroscopy, Fourier Transform Infrared/methods; beta-Lactamases/chemistry; beta-Lactamases/metabolism |
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