Blue copper proteins activation

The many redox reactions that take place within a cell make use of metalloproteins with a wide range of electron transfer potentials. To name just a few of their functions, these proteins play key roles in respiration, photosynthesis, and nitrogen fixation. Some of them simply shuttle electrons to or from enzymes that require electron transfer as part of their catalytic activity. In many other cases, a complex enzyme may incorporate its own electron transfer centers. There are three general categories of transition metal redox centers cytochromes, blue copper proteins, and iron-sulfur proteins. 

This discussion of copper-containing enzymes has focused on structure and function information for Type I blue copper proteins azurin and plastocyanin, Type III hemocyanin, and Type II superoxide dismutase s structure and mechanism of activity. Information on spectral properties for some metalloproteins and their model compounds has been included in Tables 5.2, 5.3, and 5.7. One model system for Type I copper proteins39 and one for Type II centers40 have been discussed. Many others can be found in the literature. A more complete discussion, including mechanistic detail, about hemocyanin and tyrosinase model systems has been included. Models for the blue copper oxidases laccase and ascorbate oxidases have not been discussed. Students are referred to the references listed in the reference section for discussion of some other model systems. Many more are to be found in literature searches.50... 

Copper-containing amine oxidases (non-blue copper proteins) catalyze the oxidative deamination of primary amines to the corresponding aldehydes with the release of ammonia and concomitant reduction of oxygen to hydrogen peroxide. They typically use a quinone redox cofactor [topaquinone (TPQ)], which is bound covalently in the active site, and are thought to form a Cu(I)-TPQ semi-quinone radical intermediate during the redox reaction [13]. 

Active site protonations, blue copper proteins, 36 396-398... 

Blue copper proteins, 36 323, 377-378, see also Azurin Plastocyanin active site protonations, 36 396-398 charge, 36 398-401 classification, 36 378-379 comparison with rubredoxin, 36 404 coordinated amino acid spacing, 36 399 cucumber basic protein, 36 390 electron transfer routes, 36 403-404 electron transport, 36 378 EXAFS studies, 36 390-391 functional role, 36 382-383 occurrence, 36 379-382 properties, 36 380 pseudoazurin, 36 389-390 reduction potentials, 36 393-396 self-exchange rate constants, 36 401-403 UV-VIS spectra, 36 391-393 Blue species... 

Blue copper proteins in their oxidized form contain a Cu2+ ion in the active site. The copper atom has a rather unusual tetra-hedral/trigonal pyramidal coordination formed by two histidine residues, a cysteine and a methionine residue. One of the models of plastocyanin used in our computational studies (160) is pictured in Fig. 7. Among the four proteins, the active sites differ in the distance of the sulfur atoms from the Cu center and the distortion from an approximately trigonal pyramidal to a more tetrahedral structure in the order azurin, plastocyanin, and NiR. This unusual geometrical arrangement of the active site leads to it having a number of novel electronic properties (26). 

The absorption spectra of blue copper proteins typically include one major peak and two other peaks of varying size in the range 10,000-30,000 cm-1 (164-166). MCD spectroscopy has proved useful in assigning these peaks. The electronic excitations of the active site can be classed as either d—>d or LMCT transitions. The d- fd transitions will involve excited states where the electron hole remains on the Cu atom while the LMCT transitions will move the hole to the ligands, in particular the sulfur atoms of the Met and Cys groups. Thus the d- d transitions would be expected to be more strongly influenced by spin-orbit coupling and this should be reflected in the relative size of the Cj/Dj ratios of the bands in their MCD spectra. 

Active-Site Properties of the Blue Copper Proteins A. G. Sykes... 

Metal salts with other cations are synthesized in a similar manner. Among these studies, the obtaining of coordination compounds of copper(II) in methanol with N2S2 ligand environment should be mentioned. These complexes are of permanent interest due to the modeling of active centers of blue copper proteins on their basis (see Sec. 2.2.5.4) [235-237]. Such complexes were obtained, in particular, by interaction of divalent copper perchlorate and tetrafluoroborate with very exotic ligands 661 and 662 in methanol [238] ... 

The copper proteins with a type 1 active site are commonly known as blue copper proteins due to their intense blue color in the Cu11 state. They are usually participants in electron transfer processes, and the best-known representatives of this class include plastocyanin, azurin and amicyanin [1]. The copper center in the type 1 active site is surrounded by two nitrogen donor atoms from two... 

Copper proteins are involved in a variety of biological functions, including electron transport, copper storage and many oxidase activities. A variety of reviews on this topic are available (Sykes, 1985 Chapman, 1991). Several copper proteins are easily identified by their beautiful blue colour and have been labelled blue copper proteins. The blue copper proteins can be divided into two classes, the oxidases (laccase, ascorbate oxidase, ceruloplasmin) and the electron carriers (plastocyanin, stellacyanin, umecyanin, etc.). 

The ability to exist in more than one oxidation state allows transition-metal complexes to serve as the active site of enzymes whose function is to transfer electrons (39). A great deal of effort has been directed at understanding the mechanisms of electron transfer in metalloproteins, such as cytochromes and blue copper proteins (40). Of particular interest is the mechanism by which an electron can tunnel from a metal center that is imbedded in a protein matrix to a site on the outer surface of the protein (7). A discussion of current theories is given in this volume. 

Figure 8. Proposed electron transfer pathway in blue copper proteins. The plastocyanin wave function contours have been superimposed on the blue copper (type 1) site in ascorbate oxidase (40). The contour shows the substantial electron delocalization onto the cysteine Spir orbital that activates electron transfer to the trinuclear copper cluster at 12.5 A from the blue copper site. This low-energy, intense Cys Sp - Cu charge-transfer transition provides an effective hole superexchange mechanism for rapid long-range electron transfer between these sites (2, 3, 28).

One of the major goals of studying active sites in copper proteins has therefore been to understand the spectroscopic features associated with the active site. This has led to a classification of three general types of copper protein active sites based on their unique spectral features Blue copper, normal copper and coupled binuclear copper. An additional class of copper proteins, the multi-copper oxidases, contains a combination of these three types of copper active sites. A reasonably firm understanding of the optical and EPR spectra of a number of copper proteins has now been achieved1,2K This article presents an overview of these electronic spectral features and their relationship to geometric and electronic structure. 

Type I copper is present at the active site of blue copper proteins (BCP see chapter by Nersissian and Shipp, this volume) where it is involved in the transfer of a single electron, as well as in multicopper enzymes (Gray et al., 2000 Malmstrdm, 1994 Randall et al., 2000 Sykes, 1991) (see Section V). BCP are single-domain proteins with a (3-barrel fold defined by two (3-sheets that can contain 6 to 13 strands following a Greek-key motif (Fig. 1) (Adman, 1991 Messerschmidt, 1998 Murphy ei a/., 1997 Sykes, 1991). These proteins are stable in both the reduced, Cu(I), and the oxidized, Cu(II), forms. 

B(C2H5)2The tridentate ligand [HB pz-3,5-(CH3)2 2(SCgH4-4-CH3)]" was synthesized intentionally from ArSH and K[H2B pz-3,5-(CH3)2 2]- It was converted to L MSR complexes (M = Cu or Co SR = 0-ethylcysteine,p-nitrobenzenethiolate or pentafluorophenylthiolate). These compounds were studied as synthetic approximations of the proposed active sites in the blue copper proteins (plastocyanin, azurin)... 

The active sites of oxidized blue copper proteins are characterized by unique features relative to those of normal Cu(II) complexes. These features include an intense absorption... 

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