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Literature summary for 1.3.98.3 extracted from

  • Layer, G.; Verfurth, K.; Mahlitz, E.; Jahn, D.
    Oxygen-independent coproporphyrinogen-III oxidase HemN from Escherichia coli (2002), J. Biol. Chem., 277, 34136-34142.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
hemeN gene, overexpression in Escherichia coli BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
C62S inactive mutant, no Fe-S cluster formation Escherichia coli
C66S inactive mutant, no Fe-S cluster formation Escherichia coli
C69S inactive mutant, no Fe-S cluster formation Escherichia coli
C71S inactive mutant, same Fe-S cluster formation as in wild-type HemN Escherichia coli
F68L mutant with 89% of wild-type activity Escherichia coli
G111V/G113V double mutation of Gly-111 and Gly-113, which are part of the potential GGGTP S-adenosyl-L-methionine binding motif, completely abolishes enzyme activity, reduced Fe-S cluster formation Escherichia coli
H58L inactive mutant, no Fe-S cluster formation Escherichia coli
Y56F mutant with 45% of wild-type activity and reduced Fe-S cluster formation Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
EDTA strong inhibition Escherichia coli
o-phenanthroline strong inhibition Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52000
-
gel filtration in combination with glycerol gradient centrifugation Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant HemN Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
coproporphyrinogen III + 2 S-adenosyl-L-methionine = protoporphyrinogen IX + 2 CO2 + 2 L-methionine + 2 5'-deoxyadenosine catalytic, radical mechanism Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
coproporphyrinogen-III + S-adenosyl-L-methionine HemN catalyzes the oxygen-independent conversion of coproporphyrinogen-III to protoporphyrinogen IX, requires S-adenosyl-L-methionine, NAD(P)H and additional cytoplasmatic components for catalysis. Cys-62, Cys-66 and Cys-69 are part of the conserved CXXXCXXC motif and essential for iron-sulfur cluster formation and enzyme function. Gly-111 and Gly-113 are part of the potential GGGTP S-adenosyl-L-methionine binding motif and essential for enzymatic function, catalytic, radical mechanism Escherichia coli protoporphyrinogen IX + CO2 + L-methionine + 5'-deoxyadenosine
-
?

Subunits

Subunits Comment Organism
monomer 1* 52734, sequence calculation Escherichia coli

Synonyms

Synonyms Comment Organism
More Radical SAM enzyme Escherichia coli
oxygen-independent CPO
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
4Fe-4S-center requirement, oxygen-sensitive Fe-S cluster, Cys-62, Cys-66 and Cys-69 are part of the conserved CXXXCXXC motif and essential for Fe-S cluster formation and enzyme function, Tyr-56 and His-58 are important for the Fe-S cluster integrity, His-58 may provide the fourth ligand besides the three cysteine residues Escherichia coli
NADH requires NAD(P)H, higher activity than with NADPH as cofactor Escherichia coli
NADPH requires NAD(P)H, lower activity than with NADH as cofactor Escherichia coli
S-adenosyl-L-methionine uses S-adenosyl-L-methionine as a cofactor Escherichia coli