Interconversion of two oxidized forms of taurine/alpha-ketoglutarate dioxygenase, a non-heme iron hydroxylase: evidence for bicarbonate binding.

Abstract

Taurinealpha-ketoglutarate (alphaKG) dioxygenase, or TauD, is a mononuclear non-heme iron hydroxylase that couples the oxidative decarboxylation of alphaKG to the decomposition of taurine, forming sulfite and aminoacetaldehyde. Prior studies revealed that taurine-free TauD catalyzes an O(2)- and alphaKG-dependent self-hydroxylation reaction involving Tyr-73, yielding an Fe(III)-catecholate chromophore with a lambda(max) of 550 nm. Here, a chromophore (lambda(max) 720 nm) is described and shown to arise from O(2)-dependent self-hydroxylation of TauD in the absence of alphaKG, but requiring the product succinate. A similar chromophore rapidly develops with the alternative oxidant H(2)O(2). Resonance Raman spectra indicate that the approximately 700-nm chromophore also arises from an Fe(III)-catecholate species, and site-directed mutagenesis studies again demonstrate Tyr-73 involvement. The approximately 700-nm and 550-nm species are shown to interconvert by the addition or removal of bicarbonate, consistent with the alphaKG-derived CO(2) remaining tightly bound to the oxidized metal site as bicarbonate. The relevance of the metal-bound bicarbonate in TauD to reactions of other members of this enzyme family is discussed.