Blue copper proteins binding sites

The first class is cupredoxins—single-domain blue copper proteins composed of only one BCB domain. These proteins include plastocyanin, azurin, pseudoazurin, amicyanin, auracyanins, rusticyanin, halocyanin, and sulfocyanin (see Section IV). Plantacyanin of the phytocyanin family (Section V), subunit II of the cytochrome c oxidase, and the recently characterized nitrosocyanin also fall into this class. The last two are single BCB domain polypeptides closely related structurally to cupredoxins, but harboring, respectively, a binuclear copper site known as CuA and a novel type of copper-binding site called red (see Sections IX and X). 

Intriguingly, the blue copper sites, especiaUy those with a carbonyl oxygen at the axial coordination position, display high affinity for Zn + ions. Mutants in which the Met is replaced by Gin or Glu preferentiaUy bind Zn + when expressed in heterologous systems, e.g., Escherichia coli. Examples include azurin, amicyanin, nitrite reductase, and possibly also plastocyanin (Diederix et al., 2000 Hibino et al., 1995 Murphy et al., 1995 Nar et al., 1992a Romero et al., 1993). In the case of azurin it has been shown that both wild-type and the Met—Gin mutant have the same affinity for both Zn +and Cu + (Romero ci a/., 1993). In addition, EXAFS studies showed that some preparations of blue copper proteins purihed from their natural sources also contain small fractions of Zn derivatives (DeBeer George, personal communication). 

Halocyanin and sulfocyanin are archaebacterial cupre-doxins that are attached to peripheral membranes through a lipid anchor at their N-terminus. Halocyanin was the first cupredoxin purified from an archaeon, haloalkaliphilic Natronobacterium pharaonis These cells live in high pH (around 10-11) and in extreme salinity (30%) environments. The presence of the blue copper protein, sulfocyanin in Sul-folobus acidocaldarius was first predicted from its gene sequence. It has been subsequently purified as a recombinant protein and shown to bind a single copper ion with spectroscopic properties typical for a blue copper site. ... 

A keyword search will provide access to coordinate sets from all species that are currently available as well as various site-directed mutants, type 1 copper proteins substituted with metals other than copper, and ruthenated blue copper protein derivatives. Table 1 provides a summary (with references) of the structures of blue copper-binding domains elucidated using X-ray crystallography and NMR spectroscopy. 

Electron transport between cubane [Fe4S4] clusters, cytochrome c, or Cu + centers in blue copper proteins " and the periphery of the proteins has been examined by complexing ruthenium species to surface histidines. In the case of the iron sulfur cubane in Chromatium vinosum, four surface histidines served as points of ruthenium attachment. The rates of electron transport from the Fe4S4 core to ruthenium varied over two orders of magnitude and were used to diagnose the preferred channel for electron transport. Cysteine and lysine residues have also been used as binding sites in studies of cytochrome c and cytochrome P450 cam proteins. 

Nitrite reductases catalyze both of the reactions below the physiological electron donors are either c-type cytochromes or small blue-copper proteins (eqnations 1 and 2). h28 xhe Type 1 center acts as an electron-accepting site, which then transfers the electron to the Type 2 copper where snbstrate binding and rednction occur. 

Binding of a paramagnetic, redox-inactive [Cr(CN)6]3- anion to specific sites of a blue copper protein, amicyanin, has been used in NMR-spectroscopic studies of the protein structure in solutions.285Ab initio calculations of the ligand-field spectra of [Cr(CN)6]3 have been performed and the results compared with those for cyano complexes of the other first-row transition metals.286 The role of Cr—C—N bending vibrations in the phosphorescence spectra... 

A combination of spectroscopic methods (EPR, light absorption and CD) was used in order to obtain information on the copper binding site of bovine hemocuprein (erythrocuprein), which has been considered to be intermediate in its spectroscopic properties between the blue copper proteins and complexes of copper with model peptides (94). 

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