Aldehyde, acceptor oxidase

R is an electron-donor substrate such as purine or xanthine and A is an electron acceptor such as 02 or NAD+. It is thought that the in vivo mammalian form of xanthine oxidase uses NAD+ as acceptor and is therefore, more appropriately named xanthine dehydrogenase. No evidence exists for a dehydrogenase form of aldehyde oxidase. The specificities of xanthine oxidase and aldehyde oxidase have been extensively catalogued (96), and the mechanism and properties of these enzymes have been reviewed (97, 98). 

Xanthine oxidase (XO) was the first enzyme studied from the family of enzymes now known as the molybdenum hydroxylases (HiUe 1999). XO, which catalyzes the hydroxylation of xanthine to uric acid is abundant in cow s milk and contains several cofactors, including FAD, two Fe-S centers, and a molybdenum cofactor, all of which are required for activity (Massey and Harris 1997). Purified XO has been shown to use xanthine, hypoxan-thine, and several aldehydes as substrates in the reduction of methylene blue (Booth 1938), used as an electron acceptor. Early studies also noted that cyanide was inhibitory but could only inactivate XO during preincubation, not during the reaction with xanthine (Dixon 1927). The target of cyanide inactivation was identified to be a labile sulfur atom, termed the cyanolyzable sulfur (Wahl and Rajagopalan 1982), which is also required for enzyme activity. 

The rationale for studies on flavin semiquinone metal interactions stems from the presence of flavin coenzymes which participate in electron transfer in a number of metalloflavoproteins. Iron-containing redox centers such as the heme and nonheme iron sulfur prosthetic groups (Fe2/S2, Fe+ZS, or the rubredoxin-type of iron center) constitute the more common type of metal donor-acceptor found in metalloflavoproteins, although molybdenum is encountered in the molybdenum hydroxylases (e.g. xanthine oxidase, aldehyde dehydrogenase). 

Topaquinone (TPQ). Both bacteria and eukaryotes contain amine oxidases that utilize bound copper ions and 02 as electron acceptors and form an aldehyde, NH3, and H202. The presence of an organic cofactor was suggested by the absorption spectra which was variously attributed to pyridoxal phosphate or PQQ. However, isolation from the active site of bovine serum... 

Aldehyde oxidase catalyzes the oxidation of aldehydes to carboxylic acids by dioxygen, but also catalyzes the hydroxylation of pyrimidines. Despite its rather broad specificity for substrates, it may well be that aldehyde oxidase should be regarded primarily as a pyrimidine hydroxylase. Thus, xanthine oxidase and aldehyde oxidase catalyze the hydroxylation of purines and pyrimidines respectively. The oxygen incorporated into the product comes from water, not 02. The dioxygen serves as the electron acceptor and other oxidizing agents may be used. 

Molybdenum is a component of at least three enzymes aldehyde oxidase, xanthine dehydrogenase, and sulfite oxidase. The first two contain FAD, whereas the last is a heme protein similar to cytochrome c. Xanthine dehydrogenase can also act as an oxidase, that is, it can use 02 as an electron acceptor. Physiologically, however, it uses NAD+ as an electron acceptor when it converts hypoxanthine to xanthine and the latter to uric acid (see Chapter 10). Aldehyde and sulfite oxidases are true oxidases physiologically they both use 02 as an electron acceptor. Molybdenum in all three enzymes is associated with a pterinlike cofactor whose structure is shown in Figure 6.11. The Mo cofactor cannot be... 

The deprotonation and addition of a base to thiazolium salts are combined to produce an acyl carbanion equivalent (an active aldehyde) [363, 364], which is known to play an essential role in catalysis of the thiamine diphosphate (ThDP) coenzyme [365, 366]. The active aldehyde in ThDP dependent enzymes has the ability to mediate an efScient electron transfer to various physiological electron acceptors, such as lipoamide in pyruvate dehydrogenase multienzyme complex [367], flavin adenine dinucleotide (FAD) in pyruvate oxidase [368] and Fc4S4 cluster in pyruvate ferredoxin oxidoreductase [369]. 

Krenitsky, T.A. Tuttle, J.V. Cattan, E.L. and Wang, P. A comparison of the distribution and the electron acceptor specificities of xanthine oxidase and aldehyde oxidase. Comp Biochem Physiol 49B 687-703, 1974. 

Monoamine oxidase is a flavoprotein that catalyzes the oxidative deamination of amines to form the corresponding aldehydes. 02 is the electron acceptor, and NH3 and H202 are the other products. (PNMT = phenylethanolamine-N-methyltransferase.)... 

The aliphatic alcohol oxidase, a FAD-dependent enzyme, catalyzes the oxidation of primary short-chain alcohols to the corresponding aldehydes. Dioxygen can be replaced by synthetic acceptors such as dichlorophenolindophenol or phenazine methosulfate [147l... 

Xanthine oxidase was examined for its catalytic applicability for the oxidation of aldehydes as early as 1967118]. In addition to 02, xanthine oxidase was reported to accept e.g. methylene blue, PMS or ferricyanide[19] as electron acceptors. Table 16.4-4 gives kinetic data for some substrates1201. 

XOR accelerates the hydroxylation of purines, pyrimidines, pterins and aldehydes [132]. In humans, the enzyme catalyzes the last two steps of purine catabolism the oxidation of hypoxanthine to xanthine and of the latter to uric acid. An unusual property of this, but not aU XOR enzymes [133], is its interconversion between xanthine dehydrogenase and xanthine oxidase activities which implies a switch between NAD" and molecular oxygen being used as the final electron acceptor [134]. Structural studies suggest that this switch, that can be irreversibly induced by proteolysis [135], results from conformational changes that lead to both restricted access to the NAD cofactor to its binding site and changes in the redox potential of the FAD cofactor [136],... 

The direct incomplete oxidation of sugars without phosphorylation leads to the formation of the corresponding ketones. The aldoses are oxidized into aldonic acids. The aldehydic function of this sugar is transformed into a carboxylic acid function. Glucose is oxidized into gluconic acid in this manner. The glucose oxidase catalyzes the reaction, which is coupled with the reduction of FAD. In acetic acid bacteria, electrons and protons are transported by the cytochrome chain to oxygen, which is the final acceptor. 

The flavin enzymes also are capable of transferring hydrogen (or its electrons) to acceptors other than oxygen. Methylene blue and other quinonoid dyes in in vitro experiments serve as unphysiological acceptors, whereas in vivo the soluble cytochrome c generally would seem to assume the role of the redox dye thus the formation of H2O2 is avoided. The flavoproteins, often called oxidases (e.g. amino acid oxidase, aldehyde oxidase), are facultative oxidases which usually function as dehydrogenases. 

A comparison of xanthine oxidase and aldehyde oxidase was suggested by their functional and structural similarities. Both enzymes catalyze hydroxylation reactions in which water is the source of the hydroxyl group [6,7], both have particle weights around 300,000 [8,9], and both contain FAD, molybdenum, and iron in their internal electron transport chains [7,9]. This report summarizes the results of a comparison of the distributions, substrate specificities and electron acceptor specificities of these two enzymes [10,11] and discusses the possible implications of the findings. 

Aldehyde oxidase in extracts of a wide variety of animal tissues did not usually use NAD" as an electron acceptor [11]. In contrast, with xanthine oxidase, NAD" was usually a relatively efficient electron acceptor. The various electron acceptor specificity patterns observed with xanthine oxidase using NAD", ferricyanide... 

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