Preparation of cobalt(II) derivatives of type 1 copper proteins has been extended to include bean plastocyanin and azurin from Pseudomonas aeruginosa. Fluorescence quenching data suggest that Co(II) Recently we reported the successful preparation of a Co(II) derivative (1) of a blue (type 1) (2) Clearly, an opportunity now exists for a systematic study of the Co(II) derivatives of type 1 copper proteins that have different reduction potentials. Analysis of the d-d and charge transfer bands of such Co(II) proteins conceivably could generate a reasonable structural hypothesis for the wide variation in potential of type 1 copper. Further, close comparisons of the electronic spectra of analogous Co(II) and Cu(II) proteins should allow firm assignments to be made for the intense, visible absorption bands in the latter derivatives (2).This paper is concerned with the preparation and spectroscopic study of two new Co(II) derivatives. The proteins are plastocyanin (P1) from Phaseolus vulgaris and azurin (Az) from Pseudomonas aeruginosa. Assignments are proposed for both the visible and ultraviolet absorption bands in Co(II)St, Co(II)Pl, and Co(II)Az, and a discussion of the probable nature of the low-lying electronic transitions in the native type 1 proteins is presented.
MATERIALS AND METHODSPlastocyanin was extracted from the leaves of 4-to 6-weekold bean plants and purified by the procedure of Wells (3) to an absorbance ratio A278/A597 between 1.1 and 1.2. Stellacyanin was extracted and purified as described earlier (1) LMCT, ligand-to-metal charge transfer; kK, 103 cm-'.