Y. Chen et al., Coordination of two high-affinity hexamer peptides to copper(II) and palladium(II) models of the peptide-metal chelation site on IMAC resins, INORG CHEM, 39(6), 2000, pp. 1180-1186
The coordination of peptides Ser-Pro-His-His-Gly-Gly (SPHHGG) and (His)(6)
(HHHHHH) to [Pd-II(mida)(D2O)] (mida(2-) = N-methyliminodiacetate) was stud
ied by H-1 NMR as model reactions for Cu-II(iminodiacetate)immobilized meta
l affinity chromatography (IMAC) sites. This is the first direct physical d
escription of peptide coordination for IMAC. A three-site coordination is o
bserved which involves the first, third, and fourth residues along the pept
ide chain. The presence of proline in position 2 of SPHHGG achieves the bes
t molecular mechanics and bonding angles in the coordinated peptide and enh
ances the interaction of the serine amino nitrogen. Histidine coordination
of H-1, H-3, and H-4 of (His)(6) and H-3 and H-4 of SPHHGG was detected by
H-1 NMR contact shifts and H/D exchange of histidyl protons. The EPR spectr
a of SPHHGG and HHHHHH attached to the [Cu-II(mida)] unit were obtained for
additional modeling of IMAC sites. EPR parameters of the parent [Cu(mida)(
H2O)(2)] complex are representative: g(u) = 2.31; g(yy) = 2.086; g(xx) = 2.
053; A(ll) = 161G; A(N) = 19G (three line, one N coupling). Increased rhomb
ic distortion is detected relative to the starting aqua complex in the orde
r of [Cu(mida)L] for distortion of HHHHHH > SPHHGG > (H2O)(2) The lowering
of symmetry is also seen in the decrease in the N-shf coupling, presumably
to the imino nitrogen of mida(2-) in the order 19 G (H2O), 16 G (SPHHGG) an
d 11 G (HHHHHH). Visible spectra of the [Cu(mida)(SPHHGG)] and [Cu(mida)(HH
HHHH)] as a function of pH indicate coordination of one histidyl donor at c
a. 4.5, two in the range of pH 5-7, and two chelate ring attachments involv
ing the terminal amino donor for SPHHGG or another histidyl donor of HHHHHH
in the pH domain of 7-8 in agreement with the [Pd-II(mida)L] derivatives w
hich form the two-chelate-ring attachment even at lower pH as shown by the
H-1 NMR methods.