Cobalt(II) and zinc(II) binding to a ferredoxin maquette.
We have examined the Co(II) and Zn(II) affinity of the prototype ferredoxin maquette ligand, NH(2)-KLCEGG.CIACGAC.GGW-CONH2 (IAA), which was originally designed to bind a [4Fe-4S] cluster. UV-Vis spectroscopy demonstrates tight 1:1 complex formation between Co(II) and IAA. The intensity of the S-->Co(II) charge transfer bands at 304 and 340 nm and the ligand field bands between 630 and 728 nm indicate Co(II) coordination by the four cysteine thiolates of IAA in a pseudo-tetrahedral geometry. A dissociation constant value of 5.3 microM was determined for the Co(II)-IAA complex at pH 6.5. Zn(II) readily displaces Co(II) from IAA as evinced by loss of the Co(II) spectral features. The dissociation constant for Zn(II), 20 pM at pH 6.5, was determined be competition experiments with Co(II)-IAA. These results demonstrate that the ferredoxin maquette ligand is an excellent ligand for Zn(II).
Kennedy
Michelle L
ML
Department of Chemistry, Columbia University, 3000 Broadway, MC 3121, New York, NY 10027, USA.
Petros
Amy K
AK
Gibney
Brian R
BR
D003035:Cobalt / Q000737:chemistry; D005288:Ferredoxins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008667:Metalloproteins / Q000737:chemistry; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D015032:Zinc / Q000737:chemistry
D005288:Ferredoxins; D008024:Ligands; D008667:Metalloproteins; D009842:Oligopeptides; D011994:Recombinant Proteins; D003035:Cobalt; D015032:Zinc