Product Release Rather than Chelation Determines Metal Specificity for Ferrochelatase

Amy Elizabeth Medlock, Michael Carter, Tamara A. Dailey, Harry A. Dailey, William N. Lanzilotta

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Ferrochelatase (protoheme ferrolyase, E.C. 4.99.1.1) is the terminal enzyme in heme biosynthesis and catalyzes the insertion of ferrous iron into protoporphyrin IX to form protoheme IX (heme). Within the past two years, X-ray crystallographic data obtained with human ferrochelatase have clearly shown that significant structural changes occur during catalysis that are predicted to facilitate metal insertion and product release. One unanswered question about ferrochelatase involves defining the mechanism whereby some metals, such as divalent Fe, Co, Ni, and Zn, can be used by the enzyme in vitro to produce the corresponding metalloporphyrins, while other metals, such as divalent Mn, Hg, Cd, or Pb, are inhibitors of the enzyme. Through the use of high-resolution X-ray crystallography along with characterization of metal species via their anomalous diffraction, the identity and position of Hg, Cd, Ni, or Mn in the center of enzyme-bound porphyrin macrocycle were determined. When Pb, Hg, Cd, or Ni was present in the macrocycle, the conserved π helix was in the extended, partially unwound "product release" state. Interestingly, in the structure of ferrochelatase with Mn-porphyrin bound, the π helix is not extended or unwound and is in the "substrate-bound" conformation. These findings show that at least in the cases of Mn, Pb, Cd, and Hg, metal "inhibition" of ferrochelatase is not due to the inability of the enzyme to insert the metal into the macrocycle or by binding to a second metal binding site as has been previously proposed. Rather, inhibition occurs after metal insertion and results from poor or diminished product release. Possible explanations for the lack of product release are proposed herein.

Original languageEnglish (US)
Pages (from-to)308-319
Number of pages12
JournalJournal of Molecular Biology
Volume393
Issue number2
DOIs
StatePublished - Oct 23 2009
Externally publishedYes

Fingerprint

Ferrochelatase
Metals
Heme
Porphyrins
Enzymes
Metalloporphyrins
X Ray Crystallography
Enzyme Inhibitors
Catalysis
Binding Sites
X-Rays

Keywords

  • X-ray crystallography
  • ferrochelatase
  • heme synthesis
  • metal inhibition
  • protoporphyrin IX

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Medlock, A. E., Carter, M., Dailey, T. A., Dailey, H. A., & Lanzilotta, W. N. (2009). Product Release Rather than Chelation Determines Metal Specificity for Ferrochelatase. Journal of Molecular Biology, 393(2), 308-319. https://doi.org/10.1016/j.jmb.2009.08.042

Product Release Rather than Chelation Determines Metal Specificity for Ferrochelatase. / Medlock, Amy Elizabeth; Carter, Michael; Dailey, Tamara A.; Dailey, Harry A.; Lanzilotta, William N.

In: Journal of Molecular Biology, Vol. 393, No. 2, 23.10.2009, p. 308-319.

Research output: Contribution to journalArticle

Medlock, AE, Carter, M, Dailey, TA, Dailey, HA & Lanzilotta, WN 2009, 'Product Release Rather than Chelation Determines Metal Specificity for Ferrochelatase', Journal of Molecular Biology, vol. 393, no. 2, pp. 308-319. https://doi.org/10.1016/j.jmb.2009.08.042
Medlock, Amy Elizabeth ; Carter, Michael ; Dailey, Tamara A. ; Dailey, Harry A. ; Lanzilotta, William N. / Product Release Rather than Chelation Determines Metal Specificity for Ferrochelatase. In: Journal of Molecular Biology. 2009 ; Vol. 393, No. 2. pp. 308-319.
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AB - Ferrochelatase (protoheme ferrolyase, E.C. 4.99.1.1) is the terminal enzyme in heme biosynthesis and catalyzes the insertion of ferrous iron into protoporphyrin IX to form protoheme IX (heme). Within the past two years, X-ray crystallographic data obtained with human ferrochelatase have clearly shown that significant structural changes occur during catalysis that are predicted to facilitate metal insertion and product release. One unanswered question about ferrochelatase involves defining the mechanism whereby some metals, such as divalent Fe, Co, Ni, and Zn, can be used by the enzyme in vitro to produce the corresponding metalloporphyrins, while other metals, such as divalent Mn, Hg, Cd, or Pb, are inhibitors of the enzyme. Through the use of high-resolution X-ray crystallography along with characterization of metal species via their anomalous diffraction, the identity and position of Hg, Cd, Ni, or Mn in the center of enzyme-bound porphyrin macrocycle were determined. When Pb, Hg, Cd, or Ni was present in the macrocycle, the conserved π helix was in the extended, partially unwound "product release" state. Interestingly, in the structure of ferrochelatase with Mn-porphyrin bound, the π helix is not extended or unwound and is in the "substrate-bound" conformation. These findings show that at least in the cases of Mn, Pb, Cd, and Hg, metal "inhibition" of ferrochelatase is not due to the inability of the enzyme to insert the metal into the macrocycle or by binding to a second metal binding site as has been previously proposed. Rather, inhibition occurs after metal insertion and results from poor or diminished product release. Possible explanations for the lack of product release are proposed herein.

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