Abstract
Discovery of unidentified protein functions is of biological importance because it often provides new paradigms for many research areas. Mammalian heme oxygenase (HO) enzyme catalyzes the O2-dependent degradation of heme into carbon monoxide (CO), iron, and biliverdin through numerous reaction intermediates. Here, we report that H2S, a gaseous signaling molecule, is part of a novel reaction pathway that drastically alters HO's products, reaction mechanism, and catalytic properties. Our prediction of this interplay is based on the unique reactivity of H2S with one of the HO intermediates. We found that in the presence of H2S, HO produces new linear tetrapyrroles, which we identified as isomers of sulfur-containing biliverdin (SBV), and that only H2S, but not GSH, cysteine, and polysulfides, induces SBV formation. As BV is converted to bilirubin (BR), SBV is enzymatically reduced to sulfur-containing bilirubin (SBR), which shares similar properties such as antioxidative effects with normal BR. SBR was detected in culture media of mouse macrophages, confirming the existence of this H2S-induced reaction in mammalian cells. H2S reacted specifically with a ferric verdoheme intermediate of HO, and verdoheme cleavage proceeded through an O2-independent hydrolysis-like mechanism. This change in activation mode diminished O2 dependence of the overall HO activity, circumventing the rate-limiting O2 activation of HO. We propose that H2S could largely affect O2 sensing by mammalian HO, which is supposed to relay hypoxic signals by decreasing CO output to regulate cellular functions. Moreover, the novel H2S-induced reaction identified here helps sustain HO's heme-degrading and antioxidant-generating capacity under highly hypoxic conditions.
Highlights
Discovery of unidentified protein functions is of biological importance because it often provides new paradigms for many research areas
Effects of H2S on heme oxygenase (HO) catalysis were first examined under single-turnover conditions where ferric heme complexed with rat HO-1 was converted once into products
H2S hardly affected amounts of carbon monoxide (CO) produced through the heme degradation (Fig. S4D), HPLC analysis detected two new products other than BV in the presence of H2S (Fig. 2D), whose absorption spectra are similar to each other but different from that of BV (Fig. S4A)
Summary
Discovery of unidentified protein functions is of biological importance because it often provides new paradigms for many research areas. We report that H2S, a gaseous signaling molecule, is part of a novel reaction pathway that drastically alters HO’s products, reaction mechanism, and catalytic properties. Our prediction of this interplay is based on the unique reactivity of H2S with one of the HO intermediates. HO activity could have O2 dependence that is a prerequisite to function as a primary O2 sensor in the cerebral blood vessels as well as in the carotid body [10, 12] These intricate interplays may be an inherent feature of the gaseous mediators and suggest the existence of further unidentified cross-talk. The new HO reaction, termed an S-HO reaction hereafter, is predicted on the basis of mechanistic knowledge of HO cataly-
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