Information on EC 4.98.1.1 - protoporphyrin ferrochelatase

-

Substrates: -
Products: -

?

2,4-disulfonic deuteroporphyrin + Fe2+

?

-

Substrates: -
Products: -

?

Cd2+-protoporphyrin + H+

protoporphyrin + Cd2+

-

Substrates: poor substrate
Products: -

?

Co2+-protoporphyrin + H+

protoporphyrin + Co2+

-

Substrates: -
Products: -

?

deuteroporphyrin + Fe2+

deuteroheme + 2 H+

8 entries

deuteroporphyrin + Mn2+

? + H+

705158

Substrates: in contrast to protoporphyrin IX, deuteroporphyrin lacks the vinyl groups at the 2- and 4-ring positions, and therefore, it is possible to structurally discriminate enzyme-bound (metallated) deuteroporphyrin from any possible heme (protoheme) carryover from the enzyme preparation
Products: -

?

deuteroporphyrin + Ni2+

? + H+

705158

Substrates: the Ni-deuteroporphyrin structure is distinctly different from the Mn-deuteroporphyrin model, despite the only variation in enzyme preparation being the addition of either Ni or Mn
Products: -

?

deuteroporphyrin IX + Fe2+

iron-deuteroporphyrin IX + H+

Cucumis sativus

-

653461

Substrates: -
Products: -

?

deuteroporphyrin IX + Zn2+

zinc-deuteroporphyrin IX + H+

Substrates: -
Products: -

?

deuteroporphyrin IX + Zn2+

Zn deuteroporphyrin IX + H+

-

704589

Substrates: in vitro, zinc is the preferred substrate at all concentrations of porphyrin
Products: -

?

deuteroporphyrin IX + Zn2+

Zn-deuteroporphyrin IX + H+

Fe2+-2,4-bis-acetal deuteroporphyrin

?

-

34952

Substrates: -
Products: -

?

Fe2+-2,4-diacetyldeuteroporphyrin

?

-

Substrates: -
Products: -

?

Fe2+-2,4-disulfonate deuteroporphyrin

?

-

34952

Substrates: -
Products: -

?

Fe2+-hematoporphyrin

?

Fe2+-mesoporphyrin IX + H+

mesoporphyrin IX + Fe2+

-

Substrates: -
Products: -

?

Fe2+-protoporphyrin + H+

protoporphyrin + Fe2+

-

Substrates: -
Products: -

?

hematoporphyrin + Fe2+

?

-

Substrates: -
Products: -

?

heme + Zn2+

Zn-protoporphyrin + H+

-

Substrates: -
Products: -

?

hemin + H+

mesoporphyrin IX + Zn2+

-

Substrates: -
Products: -

r

hemoglobin + Zn2+

Zn-protoporphyrin + H+

mesoporphyrin + Fe2+

mesoheme

Substrates: pH 8.0
Products: -

?

mesoporphyrin + Fe2+

mesoheme + H+

8 entries

mesoporphyrin + Zn2+

Zn-mesoporphyrin + 2 H+

mesoporphyrin IX + Co2+

cobalt mesoporphyrin IX + 2 H+

mesoporphyrin IX + Fe2+

?

mesoporphyrin IX + Fe2+

Fe-mesoporphyrin IX + 2 H+

mesoporphyrin IX + Fe2+

iron mesoporphyrin IX + 2 H+

Substrates: -
Products: -

?

mesoporphyrin IX + Fe2+

mesoheme + 2 H+

mesoporphyrin IX + ferrous ammonium sulfate

mesoheme + ammonium sulfate

Substrates: room temperature
Products: -

?

mesoporphyrin IX + Zn2+

zinc mesoporphyrin IX + 2 H+

-

34969

Substrates: -
Products: -

?

mesoporphyrin IX + Zn2+

Zn2+-mesoporphyrin + 2 H+

-

Substrates: -
Products: -

r

myoglobin + Zn2+

Zn-protoporphyrin + H+

N-methyl mesoporphyrin + Cu2+

?

666052

Substrates: pH 7.4
Products: -

?

N-methyl mesoporphyrin + Zn2+

?

666052

Substrates: pH 7.4
Products: -

?

porphyrin + Fe2+

heme + H+

Substrates: -
Products: -

?

porphyrin + metal ion

metalloporphyrin

protoheme + H+

protoporphyrin + Fe2+

protoporphyrin + Cu2+

Cu2+-protoporphyrin + H+

-

34963

Substrates: -
Products: -

?

protoporphyrin + Fe2+

protoheme + 2 H+

27 entries

protoporphyrin + Fe2+

protoheme + H+

23 entries

protoporphyrin + Zn2+

Zn-protoporphyrin + 2 H+

776152, 773858

Substrates: -
Products: -

?

protoporphyrin + Zn2+

Zn2+-protoporphyrin + H+

protoporphyrin IX + Co2+

?

Substrates: -
Products: -

?

protoporphyrin IX + Co2+

Co2+-protoporphyrin + H+

Substrates: -
Products: -

?

protoporphyrin IX + Cu2+

?

Substrates: -
Products: -

?

protoporphyrin IX + Cu2+

Cu2+-protoporphyrin + H+

Substrates: Cu2+ is a good substrate
Products: -

?

protoporphyrin IX + Fe2+

?

14 entries

protoporphyrin IX + Fe2+

Fe-protoporphyrin IX + 2 H+

protoporphyrin IX + Fe2+

Fe2+-protoporphyrin IX + 2 H+

-

763565

Substrates: -
Products: -

?

protoporphyrin IX + Fe2+

heme + 2 H+

Cucumis sativus

-

653461

Substrates: -
Products: -

?

protoporphyrin IX + Fe2+

protoheme + 2 H+

8 entries

protoporphyrin IX + Fe2+

protoheme IX + 2 H+

19 entries

protoporphyrin IX + FeCl3

protoheme + HCl

Substrates: pH 7.5, 37°C
Products: -

?

protoporphyrin IX + Ni2+

?

-

Substrates: -
Products: -

?

protoporphyrin IX + Zn2+

?

protoporphyrin IX + Zn2+

Zn protoporphyrin IX

Substrates: pH 7.4, room temperature
Products: -

?

protoporphyrin IX + Zn2+

Zn-protoporphyrin + H+

P16622

665534

Substrates: -
Products: -

?

protoporphyrin IX + Zn2+

Zn-protoporphyrin IX

protoporphyrin IX + Zn2+

Zn-protoporphyrin IX + 2 H+

Substrates: -
Products: -

?

protoporphyrin IX + Zn2+

Zn-protoporphyrin IX + H+

-

716613

Substrates: -
Products: -

?

protoporphyrin IX + Zn2+

Zn2+-protoporphyrin IX + H+

-

Substrates: -
Products: -

?

Sn2+-protoporphyrin + H+

protoporphyrin + Sn2+

-

Substrates: -
Products: -

?

Zn2+-mesoporphyrin + H+

mesoporphyrin + Zn2+

Zn2+-protoporphyrin + H+

protoporphyrin + Zn2+

additional information

?

-

24 entries

protoheme + H+

protoporphyrin + Fe2+

protoporphyrin + Fe2+

protoheme + 2 H+

protoporphyrin + Fe2+

protoheme + H+

23 entries

protoporphyrin + Zn2+

Zn-protoporphyrin + 2 H+

776152, 773858

Substrates: -
Products: -

?

protoporphyrin + Zn2+

Zn2+-protoporphyrin + H+

P42043

763563

Substrates: -
Products: -

?

protoporphyrin IX + Fe2+

?

protoporphyrin IX + Fe2+

Fe-protoporphyrin IX + 2 H+

protoporphyrin IX + Fe2+

Fe2+-protoporphyrin IX + 2 H+

-

763565

Substrates: -
Products: -

?

protoporphyrin IX + Fe2+

protoheme + 2 H+

protoporphyrin IX + Fe2+

protoheme IX + 2 H+

14 entries

protoporphyrin IX + Zn2+

Zn-protoporphyrin IX + 2 H+

Substrates: -
Products: -

?

additional information

?

-

Substrates: FECH can also catalyze the chelation between zinc and PpIX to produce zinc protoporphyrin IX (Zn-PpIX). FECH activity is estimated by the production of Zn-PpIX following the incubation of cell lysates with PpIX and zinc acetate
Products: -

-

2Fe-2S-center

-

PpfC contains a 2Fee2S cluster

heme

[2Fe-2S]-center

additional information

-

Ca2+

-

absolute requirement

Cd2+

Co2+

-

Cu2+

Fe2+

Hg2+

human ferrochelatase is capable of catalyzing the insertion of the inhibitory metal ion Hg2+ into a porphyrin macrocycle

Iron

-

Mg2+

Mn2+

Pb2+

0.1 mM, reduces the enzymatic activity by more than 60%

Sn2+

-

Zn2+

[2Fe-2S]cluster

additional information

Pseudomonas aeruginosa

-

no [2Fe-2S] cluster

2,2'-dipyridyl

-

34964

2-(4-amino-piperidin-1-yl)-ethanol

Wolbachia sp.

-

773579

-

3-benzhydryloxy-8-methyl-8-azabicyclo[3.2.1]octane

Wolbachia sp.

-

773579

-

Cd2+

Co2+

Cu2+

deferoxamine

-

deuteroporphyrin IX

-

divalent cations

-

-

EDTA

-

Fe2+

-

Zn2+-chelatase activity

34946

heme

hemin

-

Hg2+

iodacetic acid

-

iodoacetamide

ledipasvir

-

-

metalloporphyrin

-

containing Fe2+, Co2+, Zn2+, Sn2+

-

metalloporphyrins

Hordeum vulgare

-

34937

-

Mg2+

Chironomus sp.

Q967G9

0.1 mM, reduces the enzymatic activity by more than 60%

666325

Mg2+-protoporphyrin

-

34934

Mn2+

7 entries

monobromobimane

-

34952

N-alkylated porphyrins

-

N-ethylmaleimide

N-methyl mesoporphyrin

transition-state inhibitor

666052

N-methyl mesoporphyrin IX

N-methylmesoporphyrin

N-Methylprotoporphyrin

N-methylprotoporphyrin IX

-

Ni2+

-

substrate inhibition occurs when assayed in the absence of metal ion-complexing buffer components

nilotinib

-

O2

-

p-chloromercuribenzoate

-

Pb2+

porphyrin

-

high porphyrin concentrations noticeably increase the extent of zinc inhibition

704589

porphyrin isomer

-

competitive

-

Porphyrins

protohaem

-

34934

-

sulfhydryl reagents

Zn2+

additional information

13 entries

-

ascorbate

Cu2+

Chironomus sp.

Q967G9

0.1 mM, causes 4fold increase in activity

666325

L-cysteine

NaCl

additional information

-

0.056

2,4-bisacetal deuteroporphyrin

-

0.479

2,4-diacetyldeuteroporphyrin

-

0.052

2,4-disulfonic deuteroporphyrin

-

0.0052 - 0.56

Co2+

0.063 - 0.17

Cu2+

0.0213

deuteroporphyrin

-

34934

0.0026 - 0.054

deuteroporphyrin IX

0.00048 - 0.166

Fe2+

61 entries

0.055

hematoporphyrin

-

0.0057

hemin

-

pH 5.5, 45°C

0.004 - 0.1

mesoporphyrin

0.0047 - 0.0302

mesoporphyrin IX

24 entries

0.012

myoglobin

-

at pH 6.5 and 30°C

-

0.02206 - 0.1037

Ni2+

0.0121 - 0.0246

porphyrin

0.002 - 0.05

protoporphyrin

13 entries

0.00022 - 0.72

protoporphyrin IX

34 entries

0.00017 - 0.06

Zn2+

21 entries

additional information

-

0.016 - 0.033

Co2+

0.065

Cu2+

0.018 - 0.3

Fe2+

23 entries

0.52

hemin

-

pH 5.5, 45°C

6.7

mesoporphyrin IX

-

pH 8.0, 37°C

0.01167 - 0.0573

porphyrin

0.3 - 1.53

protoporphyrin

0.0125 - 1.9

protoporphyrin IX

25 entries

0.034 - 6.7

Zn2+

76 - 1200

deuteroporphyrin IX

62.5 - 416.8

Fe2+

0.000092

hemin

-

pH 5.5, 45°C

0.00101

mesoporphyrin IX

-

pH 8.0, 37°C

0.56 - 9.05

Ni2+

150 - 170

protoporphyrin

0.018 - 2015

protoporphyrin IX

0.00525 - 19000

Zn2+

0.0712 - 0.3376

deuteroporphyrin IX

0.0014 - 0.00251

N-Methylprotoporphyrin

0.007 - 2.4

Ni2+

additional information

-

Ki-values of porphyrin isomers that are not substrates are in the range of 0.013-0.07 mM, Ki-values of metalloporphyrin isomers that are not substrates are in the range of 0.002-0.013 mM, Ki-values for metal inhibitors are in the range of 0.01-0.05 mM

-

0.0015

Cyanidioschyzon merolae

-

at pH and °C

730588

0.033

R31G, pH 7.6, 30°C, unaerobic conditions

0.041

wild-type, pH 7.6, 30°C, unaerobic conditions

0.054

mutant T302A, pH 7.6, 30°C, unaerobic conditions

0.056

mutant D76G/K102T, pH 7.6, 30°C, unaerobic conditions

0.065

M11V/G104A, pH 7.6, 30°C, unaerobic conditions

0.067

mutant E61K, pH 7.6, 30°C, unaerobic conditions

0.093

E61K/L185Q/G212D, pH 7.6, 30°C, unaerobic conditions

0.413

-

0.447

-

34956

0.959

1.6

-

34944, 34953

15.2

-

pH 8.0, 37°C

3.08

pH 8.0, 25°C

4.8

presence of cysteine desulfurase NFS1 and small mitochondrial protein ISD11, pH 8.0, 25°C

747213

additional information

5.5

5.5 - 6

7.2

-

7.3

-

etioplast

34937

7.3 - 7.5

-

2 optima, pH 7.3-7.5, pH 8.0-8.2, deuteroheme formation

7.4

assay at

7.5

-

7.5 - 8

7.7

-

7.8

7.95

-

8

8 - 8.2

-

2 optima, pH 7.3-7.5, pH 8.0-8.2, deuteroheme formation

34935

8.25

-

protoporphyrin

34950

8.35

-

mesoporphyrin

34950

8.45

-

deuteroporphyrin

9

-

9.2

additional information

-

6.3 - 8.6

-

6.8 - 8.3

-

6.8 - 9.5

-

7.2 - 8.8

-

30

-

37

45

-

50

-

18 - 37

-

Q10: 2.9 kcal/mol

34931

55.8

Tm, wild-type in phosphate buffer

56

Tm, wild-type in bicine buffer

57.5

Tm, mutant E289A

61

Tm, mutant E289Q

5.12

predicted from amino acid sequence

7.35

-

isoelectric focusing

653206

Azotobacter vinelandii

-

-

-

Bdellovibrio bacteriovorus

SwissProt

-

-

-

34936, 34947, 34948, 34954, 34965, 34969

-

Bradyrhizobium japonicum

-

-

-

SwissProt

-

-

Caulobacter vibrioides

-

-

-

-

chironomid larvae

SwissProt

-

Chlamydomonas reinhardtii CC124

SwissProt

-

SwissProt

Cryptococcus neoformans

-

Cryptococcus neoformans JEC21

Q5KFH8

UniProt

-

UniProt

Cyanidioschyzon merolae

-

-

zebrafish

SwissProt

Drosophila melanogaster

-

-

Haemophilus influenzae

-

-

-

-

var. PI 177418

-

-

-

Mycobacterium tuberculosis

-

-

-

-

-

-

Neurospora crassa FGSC 4200

-

-

isoform FC1

UniProt

-

Oryza sativa Japonica Group

-

Ovis aries

-

34941

-

Phycomyces blakesleeanus

-

Phycomyces blakesleeanus NRRL1555

-

-

-

-

SwissProt

Porphyromonas gingivalis

-

Q7MXP4

SwissProt

Pseudomonas aeruginosa

-

Pseudomonas fluorescens ATCC 17400

-

Pseudomonas fluorescens

-

738261

P57778

UniProt

-

UniProt

-

-

-

34933, 34935, 34939, 34942, 34943, 34945, 34964, 729207

-

Rhizopus arrhizus

-

-

Rhizopus arrhizus RA 99-880

-

-

Shewanella loihica

Shewanella loihica PV-4

-

-

-

-

-

-

-

-

-

-

-

Uniprot

-

conversion of mesoporphyrin is dependent on the expression of ferrochelatase, since control Balb/3T3 cells have much lower expression of ferrochelatase than the ferrochelatase transfectants

-

-

-

splenic erythroblast

-

-

cultured

-

-

-

-

-

-

-

-

-

-

-

-

-

-

730585, 762872, 763565

-

-

-

-

-

additional information

apicoplast

-

mitochondrial inner membrane

parasite

-

the recombinant protein expressed in Escherichia coli

-

preprotein with a cleavable presequence at its amino-terminus

-

additional information

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

malfunction

metabolism

7 entries

physiological function

28 entries

additional information

siRNA-mediated knockdown of ferrochelatase suppresses heme synthesis and significantly increases intracellular protoporphyrin IX (PpIX) accumulation, this improves the phototoxicity of 5-aminolevulinic acid-based photodynamic therapy in urothelial cancer cell lines

706101

110000

-

1 * 110000, gel filtration in presence of sodium deoxycholate, SDS-PAGE

115000

-

1 * 115000, SDS-PAGE, gel filtration in presence of sodium deoxycholate

34944

160000

-

wild-type enzyme, gel filtration

200000

-

gel filtration

240000

-

gel filtration

250000 - 320000

-

irradiation experiments

34938

315000

-

protoporphyrin ferrochelatase PpfC, gel filtration

32500

monomer, truncated protein, SDS-PAGE

33000

Thermus thermophilus

-

x * 33000, SDS-PAGE

35000

38000

39000

-

x * 39000, SDS-PAGE

40000

41200

monomer, calculated from amino acid sequence

666325

42000

42700

-

2 * 42700, SDS-PAGE

43000

x * 43000, SDS-PAGE

45000

-

deltaH324 mutant, gel filtration

46000

-

full-length enzyme, immunoprecipitation

46100

monomer, predicted from amino acid sequence

47000

50000

-

minimum molecular weight, gel filtration in presence of Brig 35 and dithiothreitol

512000

-

complex between protoporphyrinogen dehydrogenase PgdH1 and protoporphyrin ferrochelatase PpfC, gel filtration

54430

monomer, precursor protein, predicted from amino acid sequence

60000

-

wild-type enzyme, gel filtration

63000

-

1 * 63000, SDS-PAGE

80000

84000

86000

-

gel filtration

653206

90000

mutant C406S, dimer, dynamic light scattering

additional information

?

dimer

homodimer

monomer

monomer or homodimer

-

the enzyme exists mostly as a pigment-free monomer in cells but can dimerize

763260

octamer

-

8 * 38000, SDS-PAGE, free PpfC

additional information

proteolytic modification

side-chain modification

-

lipoprotein, purified enzyme contains 0.5-1.9 mg fatty acids per mg protein

-

-

34966

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

in complex with N-methylmesoporphyrin, with and without Cu2+

652813

Mg2+ is required for the formation of Bacillus subtilis ferrochelatase crystals

overview

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

wild-type and mutants Y13F and H183C with N-methyl mesoporphyrin

-

by the hanging drop method, crystals of mutant E343K with the substrate protoporphyrin IX and mutant R115L without bound substrate. Enzyme with porphyrin bound possesses a significantly more closed active site conformation. In the substrate-bound form, the jaws of the active site mouth are closed so that the porphyrin substrate is completely engulfed in the pocket

682556

crystals of wild-type enzyme in the presence of ammonium chloride or manganese chloride, wild-type enzyme adopts a predominantly open conformation

-

681453

H263C, H341C and F337A mutants, to 2.2 A resolution

mutant enzymes E343D and E343Q in the presence of cobalt chloride, hanging drop vapor diffusion method, using 0.1 M Bis-Tris pH 6.5, 0.05 M ammonium sulfate and a range of pentaerythritol ethoxylate (15/4 EO/OH) varying from 20% to 35% (w/v)

quantum mechanical/molecular mechanics studies. The ferrous iron probably coordinates with residue Met76 at the binding site, and His263 plays the role of proton acceptor. The rate-determining step is either the first proton removed by His263 or the proton transition within the porphyrin with an energy barrier of 14.99 or 14.87 kcal/mol, respectively

748281

vapor diffusion hanging drop, 291 K, 0.05 M ammonium sulfate, 0.1 M Bis-Tris pH 6.5, 20% pentaerythritolethyloxylate, data for crystals obtained with wild-type human ferrochelatase treated with protoporphyrin IX and Hg, 1.6 A resolution; data for crystals obtained with wild-type human ferrochelatase treated with protoporphyrin IX and Cd, 1.8 A resolution, data for crystals obtained with the F110A variant of human ferrochelatase treated with deuteroporphyrin and Mn, 2.0 A resolution, data for crystals obtained with the F110A variant of human ferrochelatase treated with deuteroporphyrin and Ni, 2.20 A resolution

705158

free form, in complex with Co(II) and in complex with Cd(II) and Hg(I)

P16622

650118

in complex with beta-NAD+

-

D76G/K102T

lower KM-value than the wild-type with protoporphyrin IX as substrate, higher KM-value than the wild-type with Fe2+ as substrate

E264Q

E264V

E272S

more unstable than the wild-type, eliminates the stimulating effect of Mg2+

E61K

lower KM-value than the wild-type with protoporphyrin IX as substrate, higher KM-value than the wild-type with Fe2+ as substrate

E61K/L185Q/G212D

lower KM-value than the wild-type with protoporphyrin IX as substrate, higher KM-value than the wild-type with Fe2+ as substrate

H183A

H183C

H88A

K87A

M11V/G104A

higher KM-value than the wild-type with protoporphyrin IX as substrate, lower KM-value than the wild-type with Fe2+ as substrate

Q63A

-

reduction in enzymic activity, but not in growth rate

651702

R31G

lower KM-values than the wild-type

S54A

-

reduced growth rate, accumulation of coproporphyrin III in growth medium, but active enzyme

651702

T302A

lower KM-value than the wild-type with protoporphyrin IX as substrate, higher KM-value than the wild-type with Fe2+ as substrate

W230Y

more unstable than the wild-type, has 7% of the wild-type activity, does not display any Zn2+-induced quenching

Y13F

Y13M

E264Q

-

activity 21% of wild-type, Kd for Zn(II) is 0.0037 mM

-

E264V

-

activity less than 1% of wild-type

-

H183A

-

almost inactive enzyme, Kd for Zn(II) is 0.015 mM, 4fold decrease in affinity compared to wild-type enzyme

-

H183C

-

Kd for Zn(II) is 0.00064 mM, 6fold increased affinity compared to wild-type

-

Y13F

-

Kd for Zn(II) is 0.0048 mM, Zn(II) bound in a wild-type fashion in the structure

-

C341S

Caulobacter vibrioides

-

loss of the [2Fe-2S] cluster

H60D

-

2fold increased activity

682712

H60N

-

more than 4fold increased activity

C395delta

C406S

complements the ferrochelatase-deficient delta-hemH Escherichia coli, no activity

E343D

the catalytic efficiency of the variant is significantly diminished from the wild type enzyme

E343K

E343Q

the mutant has unmeasurable activity in vitro with iron and protoporphyrin as substrates

F110A

F110A/E343D

the catalytic efficiency of the variant is significantly diminished from the wild type enzyme

F337A

P212121 space group as the wild-type enzyme, possesses open active site pockets, spatial orientation of all side chains is identical to that of the wild-type

F417S

H157A

co-expressed with wild-type, 23-30% of wild-type activity (homogenate), loss of function (purified enzyme)

H263A

co-expressed with wild-type, 23-30% of wild-type activity (homogenate), 37.5-43% of wild-type activity (purified enzyme), exhibits a heterodimeric form when purified

H263C

P212121 space group as the wild-type enzyme, alteration in the orientation of side chains, possesses open active site pockets, has no activity

H341C

P212121 space group as the wild-type enzyme, alteration in the orientation of side chains, possesses open active site pockets, modestly decreased enzymatic activity

H386P

does not complement the ferrochelatase-deficient delta-hemH Escherichia coli, the heterodimeric form (co-expressed with wild-type) cannot be isolated as pure protein, no activity

H388A

co-expressed with wild-type, 23-30% of wild-type activity (homogenate), 37.5-43% of wild-type activity (purified enzyme), exhibits a heterodimeric form when purified

I186T

causes erythropoietic protoporphyria, co-expressed with wild-type, no activity in the homogenate

K304L

the mutant shows 7% activity compared to the wild type enzyme

M267I

causes erythropoietic protoporphyria, co-expressed with wild-type, no activity in the homogenate

M288K

does not complement the ferrochelatase-deficient delta-hemH Escherichia coli, the heterodimeric form (co-expressed with wild-type) cannot be isolated as pure protein, no activity

M76A

M76H

the mutant has a higher affinity for heme relative to the wild type enzyme

M76Y

the variant exhibits decreased kcat, as well as KM for mesoporphyrin IX and Km for iron as compared with the wild-type enzyme. The mutant is not able to utilize copper as a substrate in the in vitro

N75A

possesses intact [2Fe-2S] clusters and is as stable to storage as the wild-type

P62R

-

mutated full-length ferrochelatase, identified in a patient with erythropoietic protoporphyria, successfully targets to mitochondria and does not inhibit cleavage of the mature protein from the leader peptide

681946

Q302A

the mutant shows 40% activity compared to the wild type enzyme

Q302D

the mutant's enzyme activity is below detection limit

R115L

R115L/E343D

exhibits similar hyperbolic concentration dependencies on porphyrin and ferrous iron concentrations, characteristic two-step mechanism with rapid substrate binding followed by a slower step of metal ion chelation, 7fold decrease in kcat. Mutation primarily affects events occurring after metal ion chelation

678306

S264L

does not complement the ferrochelatase-deficient delta-hemH Escherichia coli

S303A

the mutant shows 47% activity compared to the wild type enzyme

S303P

the mutant's enzyme activity is below detection limit

S303T

the mutant's enzyme activity is below detection limit

E287A

-

low enzymic activity, protein contains protoporphyrin IX, E287 critical for control of release of product

E287Q

E289A

enhanced protein stability

E289Q

enhanced protein stability

F283L

-

the mutation eliminates inhibition by Ni2+

H207N

-

addition of metalated porphyrins yields a 1:1 complex comparable with wild type

649959

H287C

-

the mutant is substrate-inhibited by Zn2+ and Cu2+ and shows decreased inhibition by Ni2+ compare to the wild type enzyme

H287L

-

the mutant is not substrate-inhibited by Ni2+ but is weakly inhibited by Co2+, Zn2+, and inhibited by Cu2+

H287N

-

the mutant is substrate-inhibited by Ni2+, Fe2+, Zn2+, and Co2+

K250M/V251L/W256Y

K250N

-

can not form colonies onto medium containing more than 0.02 mM N-methylprotoporphyrin

P255G

-

drastically reduced inhibition of the variants by N-methylprotoporphyrin, can form colonies onto medium containing more than 0.02 mM N-methylprotoporphyrin, but is not paralleled with a decrease in specificity constant and/or catalytic activity

P255R

Q248P/S249G/K250P/G252W

S249A/K250Q/V251C

the ratio of iron/protein is 50% higher than that of the wild-type enzyme, higher KM-values than the wild-type

665512

V251L

C213S

-

loss of the [2Fe-2S] cluster, able to support the growth of the Escherichia coli deltahemH strain

C219S

-

loss of the [2Fe-2S] cluster, able to support the growth of the Escherichia coli deltahemH strain

C220S

-

loss of the [2Fe-2S] cluster, able to support the growth of the Escherichia coli deltahemH strain

C230S

-

loss of the [2Fe-2S] cluster, able to support the growth of the Escherichia coli deltahemH strain

delta209230

-

retains enough activity to complement Escherichia coli deltahemH, but is unstable after purification

D207C

Pseudomonas putida

-

does not form a [2Fe-2S] cluster

K202T/D207C

Pseudomonas putida

-

does not form a [2Fe-2S] cluster

K202T/deltaV204/D207C

Pseudomonas putida

-

does not form a [2Fe-2S] cluster

F323L

-

the mutant exhibits an enhanced Ni2+-chelatase activity over that of the parent wild type ferrochelatase

S143T

-

the mutant exhibits an enhanced Ni2+-chelatase activity over that of the parent wild type ferrochelatase

S143T/F323L

-

the mutant exhibits an enhanced Ni2+-chelatase activity over that of the parent wild type ferrochelatase

D141A

-

no enzymic activity, no dehydrogenase activity

D246V/Y248F

-

dimerization-defective, partially blocked in activity

E314A

-

no enzymic activity

G22D

-

wild type enzymic activity, no dehydrogenase activity

G47Delta

-

no enzymic activity

G47S

-

thermosensitive phenotype

34973

H235L

-

no enzymic activity

H237A

-

retains both enzymic and dehydrogenase activity

L62F

-

increase of Vmax and Km

34973

L62Y

-

no enzymic activity

S102F

-

thermosensitive phenotype

34973

FeChDELTA347

P54225

removal of the C-terminal CAB-domain led to a greatly increased turnover number, kcat, compared to the full length protein

730722

additional information

14 entries

ferrochelatase shows a calculated half-life of approximately 35 h in vivo

in absence of dithiothreitol or glycerol enzyme activity diminishes rapidly

-

influence of Sus scrofa liver drying as a stabilisation method on ferrochelatase activity for zinc protoporphyrin formation, overview. The drying process at moderate temperatures close to room conditions (10-20 C) is considered to be an effective method for FeCH preservation since it is possible to stabilise the liver, and the loss of FeCH activity is minimised, drying kinetics and mathematical model fitting

773858

loss of activity with freezing and thawing

region II is critical for both the stability and enzyme activity of Synechocystis ferrochelatase

-

716613

FECH is destabilized by oxygen and mitochondrial oxidative stress

glutathione, cysteine, dithiothreitol protect against autooxidation

-

34943

-30°C less loss of activity than under unfrozen conditions

-

34936

18°C, in 20 mM Tris-HCl (pH 8.0) containing 0.3 M NaCl, about 2 weeks, 42% residual activity

-

2 days, 4°C, 20% glycerol, 1 mM dithiothreitol, no loss of activity

-

34936

4°C, in 20 mM Tris-HCl (pH 8.0) containing 0.3 M NaCl, 1 month, no loss of activity

-

membrane-bound enzyme stable to freezing for months, storage at 4°C for at least several weeks, solubilized enzyme labile

-

34944

stable for at 1 month, -25°C, 50% loss of activity, 4°C, 1 week

-

stable for several months, pH 7.8, 70°C

-

t1/2: 12 h, 4°C or 20°C, beta-mercaptoethanol

-

34942

additional information

-

24 entries

anti-FLAG M2 affinity gel column chromatography or Ni2+-charged His-select resin column chromatography

-

716613

by gel filtration

by metal affinity chromatography

chromatography

-

complex between protoporphyrinogen dehydrogenase PgdH1 and protoporphyrin ferrochelatase PpfC, recombinant protein

-

expressed in Escherichia coli

five-step purification, by gel filtration, purified about 410fold with a 3% yield

-

immunopurification from mitochondrial extracts

P16622

665534

Ni-IMAC column chromatography and Sephacryl S-300 gel filtration

P54225

730722

Ni-NTA column chromatography. After aerobic purification, the cysteine residues can oxidize and release the [2Fe-2S] cluster, causing ferrochelatase to precipitate. CHAPS is best at solubilizing ferrochelatase, while retaining full activity as compared to the enzyme in sodium cholate

715203

Ni2+-NTA bead chromatography

Ni2+-NTA bead column chromatography, gel filtration

-

partial

precipitates immediately after purification, making it unsuitable for further analysis

Porphyromonas gingivalis

Q7MXP4

recombinant enzyme, expressed in Escherichia coli

-

recombinant protein

recombinant wild-type and mutants P255R, S249A/K250Q/V251C, K250M/V251L/W256Y, Q248P/S249G/K250P/G252W

665512

the FeCH enzyme is extracted from raw or dried pork liver, crude extract

773858

the recombinant enzyme

usage of high power ultrasound in continuous and pulsed modes to intensify the extraction of the enzyme FeCH from pork liver

776152

wild-type and deltaH324 mutant, by affinity chromatography and gel filtration

-

with Ni2+ beads

-

-

co-expression of just one native protein, ferrochelatase, in the presence of 60 microM delta-aminolevulinic acid (10 mg/l) is sufficient for 100% heme incorporation into three unrelated heme proteins derived from different organisms

Escherichia coli

-

706606

Escherichia coli deltavis cells harbouring plasmids encoding active ferrochelatase mutants, wild-type murine ferrochelatase and mutants overexpressed, under the control of the Escherichia coli alkaline phosphatase promoter phoA, in Escherichia coli BL21(DE3) cells

-

Escherichia coli expressing His-tagged ferrochelatase

-

expressed in a Saccharomyces cerevisiae hem15-deficient strain

Cryptococcus neoformans

Q5KFH8

expressed in Escherichia coli

expressed in Escherichia coli BL21 cells

-

expressed in Escherichia coli BL21(DE3) cells

expressed in Escherichia coli HB101 cells

expressed in Escherichia coli Rosetta 2 (DE3) cells

Oryza sativa Japonica Group

P54225

730722

expressed in Oryza sativa cultivar Dongjin

Q69TB1, Q0DIV0

716025

expression in Escherichia coli

expression in Escherichia coli BL21(DE3)

expression in Escherichia coli JM109

expression in Escherichia coli, wild-type and mutants E289Q and E289A

ferrochelatase protein is induced in parallel with mitoferrin-1 and Abcb10 during Friend mouse erythroleukemia cell erythroid differentiation

-

702862

into the eukaryotic expression plasmid pEYFP-N1, HeLa cells transiently transfected with YFP-tagged, wild-type FECH

-

681946

into the expression vector pTrcHisA, expressed in Escherichia coli hemH deletion mutant

into the expression vector pTrcHisA, not expressed in Escherichia coli hemH deletion mutant

Azotobacter vinelandii

-

overexpression in Escherichia coli

recombinant full-length and deltaH324 mutant expressed in Escherichia coli using the pET9a-His6 vector

-

recombinant human ferrochelatase (R115L) expressed in Escherichia coli JM109 cells

678306

wild-type and mutants expressed in Escherichia coli BL21(DE3)

ferrochelatase is light-regulated in a white collar-dependent fashion, ferrochelatase transcript levels are higher after 1 h light exposure when compared with those of the strains grown in the dark

isoform FC2 expression is most effectively stimulated by light (more than 5fold). Isoform FC1 expression is most strongly induced by cycloheximide (more than 8fold) and flagellin (5fold) treatment

-

730585

isoform HemH1 is constitutively expressed. The transcription of HemH1 is regulated by the housekeeping sigma factor RpoD and potentially regulated by OxyR

the transcription of HemH2 is regulated by the oxidative stress-associated sigma factor RpoE2

transcription of the HemH gene is mildly regulated by heme limitation. The presence of nitrate in the medium does not affect HemH expression. Compared to rich medium, minimal salt medium containing acetate, glycerol or succinate as sole sources of carbon and energy induces 1.8-3.2fold increases in HemH expression

Escherichia coli

C4ZUS9

738260

using 50 mM Tris pH 8, 3 M guanidinium hydrochloride, 0.1 M NaCl and 0.1 mM Tris (2-carboxyethyl) phosphine

analysis

biotechnology

-

co-expression with ferrochelatase along with the addition of a small amount of delta-aminolevulinic acid, is sufficient to produce fully incorporated heme protein. This method is applicable for both Cys-ligated and His-ligated heme proteins

diagnostics

drug development

food industry

Sus scrofa liver extracts with a high degree of FeCH activity are postulated as potential food ingredients for the meat industry, due to their ability to catalyse Zn-protoporphyrin (ZnPP) formation

773858

medicine

additional information