Metal substitution in a blue-copper protein: the crystal structure of cadmium-azurin at 1.8 A resolution.
Crystals of cadmium-substituted azurin have been prepared by diffusing Cd(II) into crystals of apo-azurin grown previously and their structure has been determined at high resolution by X-ray crystallography. Data to 1.8 A resolution were collected by Weissenberg photography (with image plates) using synchrotron radiation. These data were combined with a 2.2 A diffractometer data set to give 90% coverage to 1.8 A. An initial model was derived from the isomorphous Cu(II)-azurin structure, and the cadmium and ligand positions added from 'omit' maps. Refinement was by restrained least squares (program PROLSQ), to a final R value of 0.168 for all data in the range 10.0-1.8 A (23 349 reflections). The final model of 1954 protein atoms, two Cd(II) ions (occupancy 0.75), four SO(4)(2-) ions and 239 water molecules has r.m.s. deviations of 0.015, 0.045 and 0.013 A from standard bond lengths, angle distances and planar groups. The protein structure is essentially the same as that of Cu(II)-azurin, with an r.m.s. deviation of 0.18 A for 97% of main-chain atoms after superposition of the two structures. The Cd atom is within 0.2 A of the equivalent copper position, displaced slightly away from the axial Met ligand towards the carbonyl O atom of Gly45. The latter has also moved slightly towards the metal, by a rotation of the peptide unit, to give a Cd-O bond of 2.76 A. The Cd-S(Cys) bond is lengthened to 2.39 A. The coordination geometry is slightly more tetrahedral than for Cu(II), and the cadmium-oxygen interaction is consistent with the presence of an oxygen ligand in the coordination sphere of stellacyanin.
Blackwell
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Department of Chemistry and Biochemistry, Massey University, Palmerston North, New Zealand.
Andersen
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Baker
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