Advanced

Amino acid residues His183 and Glu264 in Bacillus subtilis ferrochelatase direct and facilitate the insertion of metal ion into protoporphyrin IX

Hansson, Mattias LU ; Karlberg, Tobias LU ; Rahardja, Muhammad Arys; Al-Karadaghi, Salam LU and Hansson, Mats LU (2007) In Biochemistry 46(1). p.87-94
Abstract
Ferrochelatase catalyzes the terminal step in the heme biosynthetic pathway, i.e., the incorporation of Fe(II) into protoporphyrin IX. Various biochemical and biophysical methods have been used to probe the enzyme for metal binding residues and the location of the active site. However, the location of the metal binding site and the path of the metal into the porphyrin are still disputed. Using site-directed mutagenesis on Bacillus subtilis ferrochelatase we demonstrate that exchange of the conserved residues His183 and Glu264 affects the metal affinity of the enzyme. We also present the first X-ray crystal structure of ferrochelatase with iron. Only a single iron was found in the active site, coordinated in a square pyramidal fashion by... (More)
Ferrochelatase catalyzes the terminal step in the heme biosynthetic pathway, i.e., the incorporation of Fe(II) into protoporphyrin IX. Various biochemical and biophysical methods have been used to probe the enzyme for metal binding residues and the location of the active site. However, the location of the metal binding site and the path of the metal into the porphyrin are still disputed. Using site-directed mutagenesis on Bacillus subtilis ferrochelatase we demonstrate that exchange of the conserved residues His183 and Glu264 affects the metal affinity of the enzyme. We also present the first X-ray crystal structure of ferrochelatase with iron. Only a single iron was found in the active site, coordinated in a square pyramidal fashion by two amino acid residues, His183 and Glu264, and three water molecules. This iron was not present in the structure of a His183Ala modified ferrochelatase. The results strongly suggest that the insertion of a metal ion into protoporphyrin IX by ferrochelatase occurs from a metal binding site represented by His183 and Glu264. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochemistry
volume
46
issue
1
pages
87 - 94
publisher
The American Chemical Society
external identifiers
  • wos:000243157300009
  • scopus:33846067247
ISSN
0006-2960
DOI
10.1021/bi061760a
language
English
LU publication?
yes
id
a1cf6ffb-af85-4229-8cfc-c9aaa24c87b2 (old id 680421)
date added to LUP
2007-12-12 12:00:31
date last changed
2017-02-19 03:25:13
@article{a1cf6ffb-af85-4229-8cfc-c9aaa24c87b2,
  abstract     = {Ferrochelatase catalyzes the terminal step in the heme biosynthetic pathway, i.e., the incorporation of Fe(II) into protoporphyrin IX. Various biochemical and biophysical methods have been used to probe the enzyme for metal binding residues and the location of the active site. However, the location of the metal binding site and the path of the metal into the porphyrin are still disputed. Using site-directed mutagenesis on Bacillus subtilis ferrochelatase we demonstrate that exchange of the conserved residues His183 and Glu264 affects the metal affinity of the enzyme. We also present the first X-ray crystal structure of ferrochelatase with iron. Only a single iron was found in the active site, coordinated in a square pyramidal fashion by two amino acid residues, His183 and Glu264, and three water molecules. This iron was not present in the structure of a His183Ala modified ferrochelatase. The results strongly suggest that the insertion of a metal ion into protoporphyrin IX by ferrochelatase occurs from a metal binding site represented by His183 and Glu264.},
  author       = {Hansson, Mattias and Karlberg, Tobias and Rahardja, Muhammad Arys and Al-Karadaghi, Salam and Hansson, Mats},
  issn         = {0006-2960},
  language     = {eng},
  number       = {1},
  pages        = {87--94},
  publisher    = {The American Chemical Society},
  series       = {Biochemistry},
  title        = {Amino acid residues His183 and Glu264 in Bacillus subtilis ferrochelatase direct and facilitate the insertion of metal ion into protoporphyrin IX},
  url          = {http://dx.doi.org/10.1021/bi061760a},
  volume       = {46},
  year         = {2007},
}