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Bacterial ferrochelatase turns human: Tyr13 determines the apparent metal specificity of Bacillus subtilis ferrochelatase.

Hansson, Mattias LU ; Karlberg, Tobias LU ; Söderberg, Christopher LU ; Rajan, Sreekanth LU ; Warren, Martin J; Al-Karadaghi, Salam LU ; Rigby, Stephen E J and Hansson, Mats (2011) In Journal of Biological Inorganic Chemistry 16(2). p.235-242
Abstract
Ferrochelatase catalyzes the insertion of Fe(2+) into protoporphyrin IX. The enzymatic product heme (protoheme IX) is a well-known cofactor in a wide range of proteins. The insertion of metal ions other than Fe(2+) occurs rarely in vivo, but all ferrochelatases that have been studied can insert Zn(2+) at a good rate in vitro. Co(2+), but not Cu(2+), is known to be a good substrate of the mammalian and Saccharomyces cerevisiae ferrochelatases. In contrast, Cu(2+), but not Co(2+), has been found to be a good substrate of bacterial Bacillus subtilis ferrochelatase. It is not known how ferrochelatase discriminates between different metal ion substrates. Structural analysis of B. subtilis ferrochelatase has shown that Tyr13 is an indirect... (More)
Ferrochelatase catalyzes the insertion of Fe(2+) into protoporphyrin IX. The enzymatic product heme (protoheme IX) is a well-known cofactor in a wide range of proteins. The insertion of metal ions other than Fe(2+) occurs rarely in vivo, but all ferrochelatases that have been studied can insert Zn(2+) at a good rate in vitro. Co(2+), but not Cu(2+), is known to be a good substrate of the mammalian and Saccharomyces cerevisiae ferrochelatases. In contrast, Cu(2+), but not Co(2+), has been found to be a good substrate of bacterial Bacillus subtilis ferrochelatase. It is not known how ferrochelatase discriminates between different metal ion substrates. Structural analysis of B. subtilis ferrochelatase has shown that Tyr13 is an indirect ligand of Fe(2+) and a direct ligand of a copper mesoporphyrin product. A structure-based comparison revealed that Tyr13 aligns with a Met residue in the S. cerevisiae and human ferrochelatases. Tyr13 was changed to Met in the B. subtilis enzyme by site-directed mutagenesis. Enzymatic measurements showed that the modified enzyme inserted Co(2+) at a higher rate than the wild-type B. subtilis ferrochelatase, but it had lost the ability to use Cu(2+) as a substrate. Thus, the B. subtilis Tyr13Met ferrochelatase showed the same metal specificity as that of the ferrochelatases from S. cerevisiae and human. (Less)
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organization
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Contribution to journal
publication status
published
subject
keywords
Site-Directed, Mutagenesis, Ferrochelatase/*chemistry/genetics/*metabolism, Copper/metabolism, Cobalt/metabolism, Bacillus subtilis/*enzymology, Bacterial Proteins/chemistry/genetics/metabolism, Structure-Activity Relationship, Substrate Specificity, Tyrosine/chemistry/genetics/metabolism
in
Journal of Biological Inorganic Chemistry
volume
16
issue
2
pages
235 - 242
publisher
Springer
external identifiers
  • wos:000286832200006
  • pmid:21052751
  • scopus:79951550875
ISSN
1432-1327
DOI
10.1007/s00775-010-0720-4
language
English
LU publication?
yes
id
bc592e24-a6dc-40bb-9bad-8944ab9b7931 (old id 1732289)
date added to LUP
2010-12-15 12:52:29
date last changed
2017-11-12 03:04:07
@article{bc592e24-a6dc-40bb-9bad-8944ab9b7931,
  abstract     = {Ferrochelatase catalyzes the insertion of Fe(2+) into protoporphyrin IX. The enzymatic product heme (protoheme IX) is a well-known cofactor in a wide range of proteins. The insertion of metal ions other than Fe(2+) occurs rarely in vivo, but all ferrochelatases that have been studied can insert Zn(2+) at a good rate in vitro. Co(2+), but not Cu(2+), is known to be a good substrate of the mammalian and Saccharomyces cerevisiae ferrochelatases. In contrast, Cu(2+), but not Co(2+), has been found to be a good substrate of bacterial Bacillus subtilis ferrochelatase. It is not known how ferrochelatase discriminates between different metal ion substrates. Structural analysis of B. subtilis ferrochelatase has shown that Tyr13 is an indirect ligand of Fe(2+) and a direct ligand of a copper mesoporphyrin product. A structure-based comparison revealed that Tyr13 aligns with a Met residue in the S. cerevisiae and human ferrochelatases. Tyr13 was changed to Met in the B. subtilis enzyme by site-directed mutagenesis. Enzymatic measurements showed that the modified enzyme inserted Co(2+) at a higher rate than the wild-type B. subtilis ferrochelatase, but it had lost the ability to use Cu(2+) as a substrate. Thus, the B. subtilis Tyr13Met ferrochelatase showed the same metal specificity as that of the ferrochelatases from S. cerevisiae and human.},
  author       = {Hansson, Mattias and Karlberg, Tobias and Söderberg, Christopher and Rajan, Sreekanth and Warren, Martin J and Al-Karadaghi, Salam and Rigby, Stephen E J and Hansson, Mats},
  issn         = {1432-1327},
  keyword      = {Site-Directed,Mutagenesis,Ferrochelatase/*chemistry/genetics/*metabolism,Copper/metabolism,Cobalt/metabolism,Bacillus subtilis/*enzymology,Bacterial Proteins/chemistry/genetics/metabolism,Structure-Activity Relationship,Substrate Specificity,Tyrosine/chemistry/genetics/metabolism},
  language     = {eng},
  number       = {2},
  pages        = {235--242},
  publisher    = {Springer},
  series       = {Journal of Biological Inorganic Chemistry},
  title        = {Bacterial ferrochelatase turns human: Tyr13 determines the apparent metal specificity of Bacillus subtilis ferrochelatase.},
  url          = {http://dx.doi.org/10.1007/s00775-010-0720-4},
  volume       = {16},
  year         = {2011},
}