Mammalian ferrochelatase, a new addition to the metalloenzyme family

Glória C. Ferreira, Ricardo Franco, Steven G. Lloyd, Alice S. Pereira, Isabel Moura, José J. G. Moura, Boi Hanh Huynh

Research output: Contribution to journalArticle

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Abstract

A [2Fe-2S] cluster has been detected in mammalian ferrochelatase, the terminal enzyme of the heme biosynthetic pathway. Natural ferrochelatase, purified from mouse livers, and recombinant ferrochelatase, purified from an overproducing strain of Escherichia coli, were investigated by electron paramagnetic resonance (EPR) and Mossbauer spectroscopy. In their reduced forms, both the natural and recombinant ferrochelatases exhibited an identical EPR signal with g values (g = 2.00, 1.93, and 1.90) and relaxation properties typical of [2Fe-2S]+ cluster. Mossbauer spectra of the recombinant ferrochelatase, purified from a strain of E. coli cells transformed with a plasmid encoding murine liver ferrochelatase and grown in 57Fe-enriched medium, demonstrated unambiguously that the cluster is a [2Fe-2S] cluster. No change in the cluster oxidation state was observed during catalysis. The putative protein binding site for the Fe-S cluster in mammalian ferrochelatases is absent from the sequences of the bacterial and yeast enzymes, suggesting a possible role of the [2Fe-2S] center in regulation of mammalian ferrochelatases.

Original languageEnglish
Pages (from-to)7062-7065
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number10
Publication statusPublished - 11 Mar 1994

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Ferrochelatase
Electron Spin Resonance Spectroscopy
Liver
Escherichia coli
Paramagnetic resonance
Mossbauer Spectroscopy
Biosynthetic Pathways
Mossbauer spectroscopy
Enzymes
Catalysis
Heme
Protein Binding
Yeast
Plasmids
Yeasts
Binding Sites
Oxidation

Cite this

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title = "Mammalian ferrochelatase, a new addition to the metalloenzyme family",
abstract = "A [2Fe-2S] cluster has been detected in mammalian ferrochelatase, the terminal enzyme of the heme biosynthetic pathway. Natural ferrochelatase, purified from mouse livers, and recombinant ferrochelatase, purified from an overproducing strain of Escherichia coli, were investigated by electron paramagnetic resonance (EPR) and Mossbauer spectroscopy. In their reduced forms, both the natural and recombinant ferrochelatases exhibited an identical EPR signal with g values (g = 2.00, 1.93, and 1.90) and relaxation properties typical of [2Fe-2S]+ cluster. Mossbauer spectra of the recombinant ferrochelatase, purified from a strain of E. coli cells transformed with a plasmid encoding murine liver ferrochelatase and grown in 57Fe-enriched medium, demonstrated unambiguously that the cluster is a [2Fe-2S] cluster. No change in the cluster oxidation state was observed during catalysis. The putative protein binding site for the Fe-S cluster in mammalian ferrochelatases is absent from the sequences of the bacterial and yeast enzymes, suggesting a possible role of the [2Fe-2S] center in regulation of mammalian ferrochelatases.",
author = "Ferreira, {Gl{\'o}ria C.} and Ricardo Franco and Lloyd, {Steven G.} and Pereira, {Alice S.} and Isabel Moura and Moura, {Jos{\'e} J. G.} and Huynh, {Boi Hanh}",
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Mammalian ferrochelatase, a new addition to the metalloenzyme family. / Ferreira, Glória C.; Franco, Ricardo; Lloyd, Steven G.; Pereira, Alice S.; Moura, Isabel; Moura, José J. G.; Huynh, Boi Hanh.

In: Journal of Biological Chemistry, Vol. 269, No. 10, 11.03.1994, p. 7062-7065.

Research output: Contribution to journalArticle

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T1 - Mammalian ferrochelatase, a new addition to the metalloenzyme family

AU - Ferreira, Glória C.

AU - Franco, Ricardo

AU - Lloyd, Steven G.

AU - Pereira, Alice S.

AU - Moura, Isabel

AU - Moura, José J. G.

AU - Huynh, Boi Hanh

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