Catalase HPII of Escherichia coli catalyzes the conversion of protoheme to cis-heme d

Peter C. Loewen*, Jacek Switala, Ingemar von Ossowski, Alex Hillar, Andrew Christie, Brenda Tattrie, Peter Nicholls

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Catalase HPII from aerobically grown Escherichia coli normally contains heme d but cultures grown with poor or no aeration produce HPII containing a mixture of heme d and protoheme IX. The protoheme component of HPII from anaerobically grown cells is converted into heme d during treatment of the purified enzyme with hydrogen peroxide. It is concluded that heme d found in catalase HPII is formed by the cis-hydroxylation of protoheme in a reaction catalyzed by catalase HPII using hydrogen peroxide as a substrate. The distal His128 residue of HPII is absolutely required for the protoheme to heme d conversion. Two mutant enzymes, Ala128 and Asn128, are catalytically inactive and contain only protoheme, which is unaffected by hydrogen peroxide treatment. The Asn201 residue is not an absolute requirement for heme conversion. The mutant enzyme Ala201 contains predominantly heme d and is partially active. However, insertion of a histidyl residue to give the His201 enzyme interferes with the heme conversion reaction. This mutant form is isolated as a protoheme enzyme with limited activity, and a reversible conversion to a heme d-like species occurs in vitro in the presence of continuously generated hydrogen peroxide.

Original languageEnglish
Pages (from-to)10159-10164
Number of pages6
JournalBiochemistry
Volume32
Issue number38
DOIs
Publication statusPublished - 28 Sep 1993
MoE publication typeA1 Journal article-refereed

Keywords

  • PURIFICATION
  • KATE

Cite this