Amino acid residues His183 and Glu264 in Bacillus subtilis ferrochelatase direct and facilitate the insertion of metal ion into protoporphyrin IX
(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:
https://lup.lub.lu.se/record/680421
- author
- Hansson, Mattias LU ; Karlberg, Tobias LU ; Rahardja, Muhammad Arys ; Al-Karadaghi, Salam LU and Hansson, Mats LU
- organization
- publishing date
- 2007
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Biochemistry
- volume
- 46
- issue
- 1
- pages
- 87 - 94
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000243157300009
- scopus:33846067247
- pmid:17198378
- 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
- 2016-04-01 11:38:44
- date last changed
- 2022-04-05 02:46:31
@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 (ACS)}}, 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}}, doi = {{10.1021/bi061760a}}, volume = {{46}}, year = {{2007}}, }