Redox Centers of 4-Hydroxybenzoyl-CoA Reductase, a Member of the Xanthine Oxidase Family of Molybdenum-containing Enzymes

Matthias Boll*, Georg Fuchs, Christian Meier, Alfred Trautwein, Asma El Kasmi, Stephen W. Ragsdale, Grant Buchanan, David J. Lowe

*Korrespondierende/r Autor/-in für diese Arbeit
30 Zitate (Scopus)

Abstract

4-Hydroxybenzoyl-CoA reductase (4-HBCR) is a key enzyme in the anaerobic metabolism of phenolic compounds. It catalyzes the reductive removal of the hydroxyl group from the aromatic ring yielding benzoyl-CoA and water. The subunit architecture, amino acid sequence, and the cofactor/metal content indicate that it belongs to the xanthine oxidase (XO) family of molybdenum cofactor-containing enzymes. 4-HBCR is an unusual XO family member as it catalyzes the irreversible reduction of a CoA-thioester substrate. A radical mechanism has been proposed for the enzymatic removal of phenolic hydroxyl groups. In this work we studied the spectroscopic and electrochemical properties of 4-HBCR by EPR and Mössbauer spectroscopy and identified the pterin cofactor as molybdopterin mononucleotide. In addition to two different [2Fe-2S] clusters, one FAD and one molybdenum species per monomer, we also identified a [4Fe-4S] cluster/monomer, which is unique among members of the XO family. The reduced [4Fe-4S] cluster interacted magnetically with the Mo(V) species, suggesting that the centers are in close proximity, (<15 Å apart). Additionally, reduction of the [4Fe-4S] cluster resulted in a loss of the EPR signals of the [2Fe-2S] clusters probably because of magnetic interactions between the Fe-S clusters as evidenced in power saturation studies. The Mo(V) EPR signals of 4-HBCR were typical for XO family members. Under steady-state conditions of substrate reduction, in the presence of excess dithionite, the [4Fe-4S] clusters were in the fully oxidized state while the [2Fe-2S] clusters remained reduced. The redox potentials of the redox cofactors were determined to be: [2Fe-2S]+1/+2I, -205 mV; [2Fe-2S] +1/+2 II, -255 mV; FAD/FADH/FADH, -250 mV/-470 mV; [4Fe-4S] +1/+2, -465 mV and Mo(VI)/(V)/(VI), -380 mV/-500 mV. A catalytic cycle is proposed that takes into account the common properties of molybdenum cofactor enzymes and the special one-electron chemistry of dehydroxylation of phenolic compounds.

OriginalspracheEnglisch
ZeitschriftJournal of Biological Chemistry
Jahrgang276
Ausgabenummer51
Seiten (von - bis)47853-47862
Seitenumfang10
ISSN0021-9258
DOIs
PublikationsstatusVeröffentlicht - 21.12.2001

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