Blue copper proteins as a model for investigating electron transfer processes within polypeptide matrices.
Intramolecular long-range electron transfer (ET) processes have been investigated in two types of blue copper proteins; the single-copper protein, azurin and the multi-copper oxidase, ascorbate oxidase. These have several advantages for investigating the parameters that control the above reactions: (1) Their sole physiological role is mediating or catalyzing ET processes via the evolutionary optimized copper sites. (2) The three-dimensional structures of a considerable number of blue single copper containing proteins, e.g. azurins, and of ascorbate oxidase, have been determined at high resolution. (3) These proteins have no other cofactors except for the copper ions, thus the role of the polypeptide matrix can be addressed in a more straightforward manner. In azurins, the ET from the cystine (3-26) radical-ion produced by pulse-radiolytic reduction of this single disulfide bridge, to the Cu(II) ion bound at a distance of approximately 2.6 nm has been studied, in naturally occurring and in single-site mutated azurins. The role of changing specific amino acid residues on the internal long-range electron transfer (LRET) process and its potential pathways has been investigated. It is noteworthy that this process is most probably not part of the physiological function of azurin, hence, there has not been any evolutionary selection of structural elements for the reaction. Therefore, this provides a system for an unbiased examination of structural and chemical requirements for control of this process. By contrast, in blue copper oxidases, the internal ET from the electron uptake site from substrate to the O2 reduction site is part of these enzymes catalytic cycle, presumably optimized by selective pressure. We are investigating this internal ET in ascorbate oxidase and try to resolve the relation between this enzyme's distinct functional states and the internal ET rates. We conclude that in both types of proteins, the investigated LRET proceed primarily along covalent pathways, thus providing suitable systems where the parameters controlling the efficiency of these processes can be pursued.
Farver
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O
Institute of General Chemistry, Royal Danish School of Pharmacy, Copenhagen.
Pecht
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D001204:Ascorbate Oxidase / Q000737:chemistry / Q000378:metabolism; D001400:Azurin / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D004579:Electron Transport; D046911:Macromolecular Substances; D008954:Models, Biological; D008956:Models, Chemical; D010084:Oxidation-Reduction; D011487:Protein Conformation
D001426:Bacterial Proteins; D046911:Macromolecular Substances; C017079:mauC protein, Methylobacterium extorquens; D001400:Azurin; D001204:Ascorbate Oxidase