CuA, an
electron
transfer center present in cytochrome c oxidase, COX, and
nitrous
oxide reductase, N2OR, is a dimeric copper
complex
with four ligands, two cysteine thiols bridging the metal ions and two
terminal histidine residues. The center cycles between the
mixed-valence
state [Cu(I),Cu(II)] and the reduced state [Cu(I),-Cu(I)]. The EPR,
optical
absorption, low-temperature magnetic circular dichroism, and CD spectra
of three proteins containing the mixed-valence state of CuA have been
measured
between 33 000 and 5000 cm-1. These
results
point to two forms of the chromophore, one in the enzyme N2OR
of Pseudomonas stutzeri, lacking its catalytic center, and also
in a water soluble domain of subunit II of Paracoccus denitrificans
COX and the other, referred to as CuA*, in a
site
engineered into a soluble domain of subunit II of the quinol oxidase in
Escherichia coli. An assignment of the electronic spectrum has
been
made in terms of a covalent planar core [Cu2(SR)2]+
with a Cu-S distance of 2.2 Å, a Cu-Cu distance of 2.5 Å,
and
a Cu-S-Cu angle of 70°. Molecular orbitals arising from five 3d
orbitals
on each copper and two lone-pair thiolate orbitals on each cysteine
ligand
divide into three sets, four bonding (with respect to the Cu-S
interaction)
orbitals at lower energy, four antibonding orbitals at higher energy,
and
six intermediate nonbonding orbitals. The inversion center of the
copper
core imposes rather strict selection rules giving rise to two pairs of
allowed electronic transitions, polarized along either the S-S or the
Cu-Cu
axis. A delocalization energy of ~4500 cm-1
can be estimated from the data, which is at least 1 order of magnitude
larger than the vibrational energies of the core, accounting for the
stability
of the class III or delocalized mixed-valence form. The CuA
sites in COX and N2OR have essentially identical electronic structures
with complete delocalization. However, the CuA* site shows partial
trapping
of the valences. The close approach, to ~2.3 Å, of the backbone
carbonyl
group of a conserved glutamic acid residue is proposed to be
responsible
for this partial localization. CuA is a highly
covalent
planar rhomb which provides an effective path, with low reorganization
energy, for electron transfer across subunit II from cytochrome c
to the cytochromes
a and possibly a3 of COX.