Literature summary for 4.99.1.9 extracted from

Hofbauer, S.; Helm, J.; Obinger, C.; Djinovic-Carugo, K.; Furtmueller, P.G.
Crystal structures and calorimetry reveal catalytically relevant binding mode of coproporphyrin and coproheme in coproporphyrin ferrochelatase (2020), FEBS J., 287, 2779-2796.

Cloned(Commentary)

Cloned (Comment)	Organism
gene cpfC, sequence comparisons and phylogenetic analysis, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli	Listeria monocytogenes

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme in apoform and with bound product coproheme, mixing of 9.5 mg/ml protein with or without bound ligand in solution with 16% w/v PEG 8000, 20% v/v glycerol, and 0.04 M KH2PO4 for the apo-enzyme crystals and with 0.1 M Bis-Tris, pH 5.5, 25% PEG 3350, 0.2 M MgCl2 for the complexed enzyme, 2 days, X-ray diffraction structure determination and analysis, molecular replacement using the PDB structure 2HK6 of BsCpfC enzyme as template	Listeria monocytogenes

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme in apoform and with bound product coproheme, mixing of 9.5 mg/ml protein with or without bound ligand in solution with 16% w/v PEG 8000, 20% v/v glycerol, and 0.04 M KH2PO4 for the apo-enzyme crystals and with 0.1 M Bis-Tris, pH 5.5, 25% PEG 3350, 0.2 M MgCl2 for the complexed enzyme, 2 days, X-ray diffraction structure determination and analysis, molecular replacement using the PDB structure 2HK6 of BsCpfC enzyme as template

Listeria monocytogenes

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics	Listeria monocytogenes
0.00028	-	Fe2+	pH 7.0, 25°C, recombinant enzyme	Listeria monocytogenes

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
coproporphyrin III + Fe2+	Listeria monocytogenes	-	Fe-coproporphyrin III + 2 H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Listeria monocytogenes	A0A5D5VUB4	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and desalting gel filtration	Listeria monocytogenes

Renatured (Commentary)

Renatured (Comment)	Organism
reconstitution of LmCpfC for crystallization with coproheme is performed by the addition of a 1.2 fold molar excess, and the complex is purified by gel filtration	Listeria monocytogenes

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
coproporphyrin III + Fe2+	-	Listeria monocytogenes	Fe-coproporphyrin III + 2 H+	-	?
coproporphyrin III + Fe2+	the reaction is carried out anaerobically under argon flow and in the dark	Listeria monocytogenes	Fe-coproporphyrin III + 2 H+	-	?
additional information	when LmCpfC is incubated with both substrates, Fe2+ and coproporphyrin III, Fe-coproporphyrin III (coproheme) is the only detected species	Listeria monocytogenes	?	-	-

Synonyms

Synonyms	Comment	Organism
CpfC	-	Listeria monocytogenes
ferrochelatase	-	Listeria monocytogenes
LmCpfC	-	Listeria monocytogenes

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Listeria monocytogenes

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	binding of coproporphyrin III or coproheme to apo-LmCpfC has a strong stabilizing effect on the protein, as reflected in the increased thermal stability of the resulting complexes	Listeria monocytogenes

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.132	-	Fe2+	pH 7.0, 25°C, recombinant enzyme	Listeria monocytogenes

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Listeria monocytogenes

General Information

General Information	Comment	Organism
evolution	phylogenetic analysis	Listeria monocytogenes
metabolism	coproporphyrin ferrochelatases (CpfCs) insert ferrous iron into coproporphyrin III yielding coproheme. CpfCs are utilized by prokaryotic, mainly monoderm (Gram-positive) bacteria within the coproporphyrin-dependent (CPD) heme biosynthesis pathway	Listeria monocytogenes
additional information	analysis of the catalytically relevant binding mode of coproporphyrin and coproheme in coproporphyrin ferrochelatase, crystal structures analysis and calorimetric methods, detailed overview. UV-vis absorption spectroscopy of LmCpfC with the substrate coproporphyrin III and the product coproheme, spectral transitions upon binding of coproporphyrin III to apo-LmCpfC shows a biphasic behavior, structure comparisons. Binding of coproporphyrin III or coproheme to apo-LmCpfC has a strong stabilizing effect on the protein, as reflected in the increased thermal stability of the resulting complexes	Listeria monocytogenes
physiological function	coproporphyrin ferrochelatases (CpfCs) insert ferrous iron into coproporphyrin III yielding coproheme. CpfCs are utilized by prokaryotic, mainly monoderm (Gram-positive) bacteria within the coproporphyrin-dependent (CPD) heme biosynthesis pathway	Listeria monocytogenes