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Updated: Sep 29, 2026

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
Published on: December 16, 2013
A new type 2 copper cysteinate azurin. Involvement of an engineered exposed cysteine in copper binding through
Irene M C van Amsterdam1, Marcellus Ubbink, Marieke van den Bosch
1Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, Einsteinweg 55, P. O. Box 9502, The Netherlands.
Abstract:
The double mutant H117G/N42C azurin exhibits tetragonal type 2 copper site characteristics with Cys(42) as one of the copper ligands as concluded from spectroscopic evidence (UV-visible, EPR, and resonance Raman). Analysis of the kinetics of copper uptake by the apoprotein by means of stopped flow spectroscopy suggests that the solvent-exposed Cys(42) assists in binding the metal ion and carrying it over to the active site where it becomes coordinated by, among others, a second cysteine, Cys(112). A structure is proposed in which the loop from residue 36 to 47 has rearranged to form a tetragonal type 2 copper site with Cys(42) as one of the ligands. The process of copper uptake as observed for the double mutant may be relevant for a better understanding of the way copper chaperones accept and transfer metal ions in the living cell.
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