N. Bonander et al., THE METAL SITE OF PSEUDOMONAS-AERUGINOSA AZURIN, REVEALED BY A CRYSTAL-STRUCTURE DETERMINATION OF THE CO(II) DERIVATIVE AND CO-EPR SPECTROSCOPY, Proteins, 27(3), 1997, pp. 385-394
The crystal structure of cobalt-substituted azurin from Pseudomonas ae
ruginosa has been determined to final crystallographic R value of 0.17
5 at 1.9 Angstrom resolution, There are four molecules in the asymmetr
ic unit in the structure, and these four molecules are packed as a dim
er of dimers, The dimer packing is very similar to that of the wild-ty
pe Pseudomonas aeruginosa azurin dimer. Replacement of the native copp
er by the cobalt ion has only small effects on the metal binding site
presumably because of the existence of an extensive network of hydroge
n bonds in its immediate neighborhood, Some differences are obvious, h
owever, In wild-type azurin the copper atom occupies a distorted trigo
nal bipyramidal site, while cobalt similar to zinc and nickel occupy a
distorted tetrahedral site, in which the distance to the Met121,S-del
ta atom is increased to 3.3-3.5 Angstrom and the distance to the carbo
nyl oxygen of Gly45 has decreased to 2.1-2.4 Angstrom, The X-band EPR
spectrum of the high-spin Co(II) in azurin is well resolved (apparent
g values g(x)' = 5.23; g(y)' = 3.83; g(z)' = 1.995, and hyperfine spli
ttings A(x)' = 31; A(y)' = 20-30; A(z)' = 53 G) and indicates that the
ligand field is close to axial. (C) 1997 Wiley-Liss, Inc.