Biochemical Journal: this is an Accepted Manuscript, not the final Version of Record. You are
encouraged to use the Version of Record that, when published, will replace this version. The most
up-to-date version is available at http://dx.doi.org/10.1042/BCJ20160696. Please cite using the DOI
10.1042/BCJ20160696
BIOCHEMICAL JOURNAL
ACCEPTED MANUSCRIPT
The HemQ coprohaem decarboxylase generates reactive oxygen
species: implications for the evolution of classical haem biosynthesis
Charlie Hobbs, Harry A. Dailey and Mark Shepherd
Cite as Biochemical Journal (2016) DOI: 10.1042/BCJ20160696
Copyright 2016 The Author(s).
Use of open access articles is permitted based on the terms of the specific Creative Commons Licence under which the
article is published. Archiving of non-open access articles is permitted in accordance with the Archiving Policy of Portland Press
(http://www.portlandpresspublishing.com/content/open-access-policy#Archiving).
Use of open access articles is permitted based on the terms of the specific Creative Commons Licence under
which the article is published. Archiving of non-open access articles is permitted in accordance with the
Archiving Policy of Portland Press (
http://www.portlandpresspublishing.com/content/open-access-policy#Archiving).
ACCEPTED MANUSCRIPT
Bacteria require a haem biosynthetic pathway for the assembly of a variety of protein complexes
including cytochromes, peroxidases, globins, and catalase. Haem is synthesised via a series of
tetrapyrrole intermediates including non-metallated porphyrins such as protoporphyrin IX, which is
well-known to generate reactive oxygen species (ROS) in the presence of light and
oxygen. Staphylococcus aureus has an ancient haem biosynthetic pathway that proceeds via the
formation of coproporphyrin III, a less reactive porphyrin. Herein, we demonstrate for the first time
that HemY of S. aureus is able to generate both protoporphyrin IX and coproporphyrin III, and that
the terminal enzyme of this pathway, HemQ, can stimulate the generation of protoporphyrin IX (but
not coproporphyrin III). Assays with hydrogen peroxide, horseradish peroxidase, superoxide
dismutase, and catalase confirm that this stimulatory effect is mediated by superoxide. Structural
modelling reveals that HemQ enzymes do not possess the structural attributes that are common to
peroxidases that form compound I [FeIV=O]+, which taken together with the superoxide data leaves
Fenton chemistry as a likely route for the superoxide-mediated stimulation of protoporphyrinogen
IX oxidase activity of HemY. This generation of toxic free radicals could explain why HemQ enzymes
have not been identified in organisms that synthesise haem via the classical protoporphyrin IX
pathway. This work has implications for the divergent evolution of haem biosynthesis in ancestral
microorganisms and provides new structural and mechanistic insights into a recently discovered
oxidative decarboxylase reaction.
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