Divalent copper was found to inhibit non-competitively the lysis of Micrococcus lysodeikticus cells by hen egg-white lysozyme, with an inhibition constant Ka= 3.8 × 102m-1. The association constants of Cu2+ for lysozyme and for a derivative of lysozyme in which tryptophan residue 108 was selectively modified, were measured spectrofluorimetrieally and found to be 1.8 × 102 m-1 and 1.0 × 103m-1, respectively. The electron spin resonance spectrum of Cu2+ was not affected by the addition of lysozyme, whereas many new lines appeared on addition of the modified protein. This was interpreted as evidence for the binding of Cu2+ in the neighbourhood of tryptophan 108. To unequivocally establish the site of ligation of Cu2+, crystals of lysozyme soaked in Cu2+ were examined by X-ray crystallography and the results compared to those obtained from crystals of native lysozyme. Cu2+ was found to be located 2 to 3 Å from the carboxyl side-chain of aspartic acid 52, 5 Å from the carboxyl of glutamic acid 35 and about 7 Å from tryptophan 108. The addition of a saccharide inhibitor to lysozyme was found to increase the association constant of Cu2+ for lysozyme from a value of 1.8 × 102 m-1 to 6.0 × 102 m-1. This finding was interpreted as indicative of a change in conformation around tryptophan 108 and glutamic acid 35 induced by the interaction of saccharides with the enzyme, which affects the metal binding properties of aspartic acid 52. © 1974.
Teichberg, V. I., Sharon, N., Moult, J., Smilansky, A., & Yonath, A. (1974). Binding of divalent copper ions to aspartic acid residue 52 in hen egg-white lysozyme. Journal of Molecular Biology, 87(2), 357–368. https://doi.org/10.1016/0022-2836(74)90155-7