Cloned (Comment) | Organism |
---|---|
wild-type and mutants expressed in Escherichia coli BL21(DE3) | Bacillus subtilis |
Crystallization (Comment) | Organism |
---|---|
crystals of wild-type and mutants H183A, H88A and K87A with iron, by addition of solid (NH4)2Fe(SO4)2 x 6 H2O, 1.2 A to 1.7 A resolution | Bacillus subtilis |
structure in presence of iron. Only a single iron ion is 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 ion is not present in the structure of a His183Ala modified ferrochelatase. Insertion of a metal ion into protoporphyrin IX by ferrochelatase occurs from a metal binding site represented by His183 and Glu264 | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
E264Q | 21% of wild-type activity | Bacillus subtilis |
E264Q | activity 21% of wild-type, Kd for Zn(II) is 0.0037 mM | Bacillus subtilis |
E264V | less than 1% of wild-type activity | Bacillus subtilis |
E264V | activity less than 1% of wild-type | Bacillus subtilis |
H183A | less than 1% of wild-type activity | Bacillus subtilis |
H183A | almost inactive enzyme, Kd for Zn(II) is 0.015 mM, 4fold decrease in affinity compared to wild-type enzyme | Bacillus subtilis |
H183C | less than 1% of wild-type activity | Bacillus subtilis |
H183C | Kd for Zn(II) is 0.00064 mM, 6fold increased affinity compared to wild-type | Bacillus subtilis |
H88A | 5% of wild-type activity | Bacillus subtilis |
H88A | Kd for Zn(II) is 0.0028 mM, activity is 5% of wild-type | Bacillus subtilis |
K87A | 92% of wild-type activity | Bacillus subtilis |
K87A | retains 92% of wild-type activity, Kd for Zn(II) is 0.013 mM | Bacillus subtilis |
Y13F | 71% of wild-type activity | Bacillus subtilis |
Y13F | Kd for Zn(II) is 0.0048 mM, Zn(II) bound in a wild-type fashion in the structure | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | single iron in the active site, coordinated in a square pyramidal fashion by two amino acid residues, His183 and Glu264, and three water molecules | Bacillus subtilis | |
Mg2+ | - |
Bacillus subtilis | |
Zn2+ | - |
Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | P32396 | - |
- |
Bacillus subtilis 168 | P32396 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
protoporphyrin IX + Fe2+ | insertion of a metal ion into protoporphyrin IX by ferrochelatase occurs from a metal binding site represented by His183 and Glu264 | Bacillus subtilis | ? | - |
? | |
protoporphyrin IX + Fe2+ | insertion of a metal ion into protoporphyrin IX by ferrochelatase occurs from a metal binding site represented by His183 and Glu264 | Bacillus subtilis 168 | ? | - |
? |