Information on EC 2.5.1.61 - hydroxymethylbilane synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

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EC NUMBER
COMMENTARY hide
2.5.1.61
-
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RECOMMENDED NAME
GeneOntology No.
hydroxymethylbilane synthase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
4 porphobilinogen + H2O = hydroxymethylbilane + 4 NH3
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
of NH3, C-N bond cleavage
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
tetrapyrrole biosynthesis I (from glutamate)
-
-
tetrapyrrole biosynthesis II (from glycine)
-
-
heme metabolism
-
-
Porphyrin and chlorophyll metabolism
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
Microbial metabolism in diverse environments
-
-
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SYSTEMATIC NAME
IUBMB Comments
porphobilinogen:(4-[2-carboxyethyl]-3-[carboxymethyl]pyrrol-2-yl)methyltransferase (hydrolysing)
The enzyme works by stepwise addition of pyrrolylmethyl groups until a hexapyrrole is present at the active centre. The terminal tetrapyrrole is then hydrolysed to yield the product, leaving a cysteine-bound dipyrrole on which assembly continues. In the presence of a second enzyme, EC 4.2.1.75 uroporphyrinogen-III synthase, which is often called cosynthase, the product is cyclized to form uroporphyrinogen-III. If EC 4.2.1.75 is absent, the hydroxymethylbilane cyclizes spontaneously to form uroporphyrinogen I.
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CAS REGISTRY NUMBER
COMMENTARY hide
9036-47-9
-
9074-91-3
-
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
putative
UniProt
Manually annotated by BRENDA team
gene hemC
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
sulfate-reducing bacterium
-
-
Manually annotated by BRENDA team
TG1
-
-
Manually annotated by BRENDA team
wild type siblings of mutant cats or unrelated wild type cats, porphyric mutant cats type Saskatchewan (c.189dupT = trunkated (premature stop codon 65) L64S), type Massachusetts (c842_844delGAG = delGly281), type Missouris (c.445C>T = R149W), type OregonAR (c250G>A = A84T)
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
soya-bean
-
-
Manually annotated by BRENDA team
pea
-
-
Manually annotated by BRENDA team
Sprague-Dawley
-
-
Manually annotated by BRENDA team
wild-type D3, Sammlung von Algenkulturen, Pflanzenphysiologisches Institiut Universit„t G”ttingen, Germany
-
-
Manually annotated by BRENDA team
maize
-
-
Manually annotated by BRENDA team
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
molecular dynamics simulations for determmination of the exit mechanism of hydroxymethylbilane from the enzyme at the end of the catalytic cycle
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4 porphobilinogen + H2O
hydroxymethylbilane + 4 NH3
show the reaction diagram
porphobilinogen
?
show the reaction diagram
porphobilinogen + H2O
1-hydroxymethylbilane + 4 NH3
show the reaction diagram
porphobilinogen + H2O
hydroxylmethylbilane + NH3
show the reaction diagram
porphobilinogen + H2O
hydroxymethylbilane + 4 NH3
show the reaction diagram
-
polymerization of 4 pyrrole molecules to a linear tetrapyrrole
i.e. pre-uroporphyrinogen, highly unstable compound
-
?
porphobilinogen + H2O
hydroxymethylbilane + NH3
show the reaction diagram
porphobilinogen + H2O
uroporphyrinogen III
show the reaction diagram
additional information
?
-
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4 porphobilinogen + H2O
hydroxymethylbilane + 4 NH3
show the reaction diagram
porphobilinogen
?
show the reaction diagram
porphobilinogen + H2O
1-hydroxymethylbilane + 4 NH3
show the reaction diagram
-
third reaction step in heme biosynthesis pathway
-
-
?
porphobilinogen + H2O
hydroxymethylbilane + NH3
show the reaction diagram
additional information
?
-
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dipyrromethane
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-bromoporphobilinogen
-
-
2-methylopsopyrroledicarboxylic acid
-
-
5,5'-dithiobis(2-nitrobenzoic acid)
-
5 mM, pH 8.0, 50% inhibition
Ag2+
-
1 mM AgCl2, 90% inhibition
Coproporphyrinogen
-
-
Cr3+
-
strong
glycerol
-
15% inhibits 40% of enzyme activity
hydroxylamine
isoporphobilinogen
-
-
K+
-
at high concentrations
methoxyamine
-
-
N-ethylmaleimide
NaBH4
-
partially inactivates
opsopyrroledicarboxylic acid
p-chloromercuribenzoate
p-hydroxymercuribenzoate
Pb2+
-
strong
PbNO3
-
0.005 mM, 35% inhibition
-
protoporphyrin IX
-
enzyme activity decreases with increasing concentrations, reaching a 33% inhibition at the vivo concentration of porphyrin, 0.00185 mM, and 0.014 mM porphobilinogen
protoporphyrinogen
-
inhibits both erythroid and lymphoblast forms of the enzyme
pyridoxal 5'-phosphate
-
partially inactivates
additional information
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.013 - 0.085
Hydroxymethylbilane
0.0011 - 1.579
porphobilinogen
additional information
additional information
-
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.25
porphobilinogen
-
-
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000039
-
purified recombinant GST-tagged mutant enzyme
0.001
mutant enzyme D44A, at 37°C
0.00117
-
-
0.002
-
R167Q mutant, pH 8.0
0.0128
-
40 kDa enzyme form
0.0183
-
42 kDa enzyme form
0.022
-
purified recombinant GST-tagged wild-type enzyme
0.023
-
wild type, pH 8.0
0.038
-
A form
0.039
-
B form
0.04
-
-
0.0402
-
-
0.0813
-
-
0.113
-
at pH 7.4
0.333
-
200 nM porphobilinogen, 50 mM Tris-HCl, pH 8.0, 25°C
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2 - 7.3
-
-
7.5
-
at 37°C
7.8
-
mutant enzyme T59I
7.8 - 8
-
0.1-0.4 mM porphobilinogen
7.9 - 8.2
-
-
additional information
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
-
pH 5.0: unstable below, pH 9.0 stable above
6 - 10
-
pH 6.0: about 15% of maximal activity, pH 10.0: 25% of maximal activity
6 - 10.6
-
no activity below pH 6.0 and above pH 10.6
6 - 8
-
pH 6.0: enzyme is rapidly and irreversibly inactivated below, pH 8.0: about 50% of maximal activity
6.2 - 8.4
-
pH 6.2: 50% of maximal activity, pH 8.4: 30% of maximal activity
6.3 - 7.5
-
pH 6.3 and pH 7.5: 85% of maximal activity
6.5 - 9
-
pH 6.5: 25% of maximal activity, pH 9.0: 50% of maximal activity
6.5 - 9.2
-
50% of maximal activity at pH 6.5 and pH 9.2
additional information
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
activity twice that at 37°C
60
-
maximal activity
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 80
-
10% of maximal activity at 20°C, 8% of maximal activity at 80°C
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
in developing ears
Manually annotated by BRENDA team
-
seedling roots
Manually annotated by BRENDA team
-
in developing tassels
Manually annotated by BRENDA team
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25000
-
gel filtration
31000
-
gel filtration
34000
-
1 * 34000, calculation from crystal structure, asymmetric unit
34268
-
1 * 34268, calculation from sequence of amino acid
35000 - 36000
-
gel filtration
35500
-
gel filtration
36927
-
1 * 36927, calculation from sequence of cDNA
37000
-
1 * 37000, SDS-PAGE, A and B form
39000
-
sucrose density gradient centrifugation
39100
-
1 * 39100, SDS-PAGE
39500
-
1 * 39500, SDS-PAGE, B2 and B3
40500
erythroid enzyme, SDS-PAGE
41200
-
1 * 41200, SDS-PAGE, erythrocyte enzyme
44000
-
gel filtration
53000
-
GST-tagged recombinant p.Ala226ProfsX28, removing the GST tag produces a strongly degraded enzyme
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme with bound cofactor, crystallization in the dark due to light-sensitivity of the cofactor, hanging drop method, mixing of 5 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 5 mM DTT, with reservoir solution containing 25% w/v PEG 4000, 100 mM sodium citrate, pH 5.6, and 200 mM ammonium sulfate, X-ray diffraction structure determination and analysis at 1.45 A resolution, molecular modelling
purified recombinant detagged enzyme, mixing of 2.5 mg/ml protein with 0.1 M sodium cacodylate, pH 6.5-6.8, 0.2 M magnesium acetate, and 25-30%PEG 8000, room temperature, removal of the His tag is necessary to obtain enzyme crystals, X-ray diffraction structure determination and analysis at 1.46-1.60 A resolution
-
purified recombinant detagged enzyme, mixing of 2.5 mg/ml protein with 0.1 M sodium cacodylate, pH 6.5-6.8, 0.2 M magnesium acetate, and 25-30%PEG 8000, X-ray diffraction structure determination and analysis at 1.46-1.60 A resolution
-
SeMet-labelled enzyme
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vapor diffusion method, hanging drops with 20 mM Tris-HCl buffer, pH 8.2, containing 5 mM dithiothreitol and reservoir solution consisting of 0.6 M ammonium sulfate, 1.2 M lithium sulfate, 5% ethylene glycol, 50 mM sodium citrate, pH 5.6, and 50 mM dithiothreitol, diffraction data are collected at -173°C in 30% glycerol cryoprotected protein crystals
-
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9
-
stable between
489929
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 80
the enzyme loses very little activity even when heated to 80°C and then assayed at 37°C
50
A84T mutant houskeeping enzyme (OregonAR cat) at pH 8.0 has a prolonged half-life of 6.9 h compared to 1.9 h for the wild-type; A84T mutant houskeeping enzyme (OregonAR cat) at pH 8.0 has a prolonged half-life of 6.9 h compared to 1.9 h for the wild-type
56
-
2h, no loss of activity
75
-
13% loss of activity after 1h
80
-
10 min, 40% loss of activity
additional information
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
DTT stabilizes
-
enzyme is most stable in 0.3 M LiBr, 0.1 M 2-mercaptoethanol, 30% glycerol, 0.15 M Tris chloride, pH 8.0 to 9.0, or 0.3 M (NH4)2SO4
-
phosphate stabilizes
-
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
photooxidation of the enzyme in the presence of methylene blue, 0.03%, and roes bengal, 0.03%, over a 5 min period, inhibits 90% and 60% of the enzyme activity, respectively
-
5883
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15°C, purified enzyme rapidely loses its activity
-
-20°C or 4°C, unstable, loss of activity after several days
-
-20°C, crude plastide lysate is stable for at least 2 weeks
-
-20°C, forms A to E, stable for up to 18 months in 10 mM potassium phosphate buffer, pH 8.0, containing 0.2 mM dithioerythritol
-
-20°C, stable for over 1 year
-
-80°C, 50 mM Tris buffer, pH 7.5, 5% glycerol, and 5 mM beta-mercaptoethanol, 1 year, remains stable
0-4°C, less than 5% loss of activity over 10 days
-
0°C, stable in solution at pH 7.5 and 9.5 for 3 weeks
-
4°C, 50 mM Tris buffer, pH 7.5, 5% glycerol, and 5 mM beta-mercaptoethanol, 1 week, without significant change of activity
4°C, purified enzyme stable for more than a month
-
erythrocytes lysates stored in 1 mM potassium phosphate, pH 7.6, containing 0.05% Trition X-100, 1 mM dithiothreitol and 1 mM MgCl2, no or little loss of activity after 1 year
-
purified enzyme loses its activity when it is frozen
-
stored in 0.1 M Tris chloride, pH 8.5, containing 30% glycerol and 0.01 M 2-mercaptoethanol. Under these conditions 50% loss of activity after 3 weeks
-
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 forms
-
3 isoenzymes
-
5 forms, A: native enzyme, B-E: isomeres corresponding to the enzyme-substrate intermediates
-
blood samples are washed in phosphate buffered saline, liver rinsed in phosphate buffered saline
cell harvesting by centrifugation, washed, resuspended in NaCl-phosphate buffer containing of 140 mN NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3 with protease inhibitor cocktail, and 0.5% Triton X-100, lysis by shaking with lysozyme on ice, sonication, centrifugation, supernatant loaded onto glutathione sepharose 4B column, washed with 20 mM Tris-HCl, 100 mM NaCl, 1 mM EDTA, 0.5% Nonidet P-40+, pH 8.2, proteins eluted in 50 mM Tris-HCl, pH 8.3 buffer with 20 mM glutathione, thrombin digestion with thrombin at 20 U/mg protein at 20°C overnight, addition of glycerol to a concentration of 20%
-
cells centrifuged, washed with 0.9 M NaCl, re-centrifuged, washed with 20 mM Tris-HCl buffer, pH 8.2, with 5 mM dithiothreitol, and 200 microM PMSF, sonication, heating to 60°C under N2 gas, cooled to 4°C, supernatant applied to a Pharmacia 50 K/30 column packed with DEAE-Sephacel anion-exchange resin, equilibrated with 50 mM Tris-HCl buffer, pH 8.2 containing 5 mM dithiothreitol, and 100 microM PMSF, elution with 0-70 mM KCl gradient, active fractions are pooled and concentrated by ultrafiltration with a PM-10 membrane, further concentrated with a Centricon YM-10 centrifugal filter and gel-filtered on a Hiload 16/60 Superdex G-75 column, equilibrated with 100 mM Tris-HCl buffer, pH 8.2, containing 5 mM dithiothreitol and 100 microM PMSF, concentrated with a Centricon YM-10, buffer-exchanged into 20 mM Tris-HCl buffer, pH 8.2, containing 5 mM dithiothreitol with a Pharmacia PD10 column
-
cells harvested and resuspended in 20 mM Tris-HCl buffer, pH 8.0 containing 500 mM NaCl and 10 mM imidazole, cells are lysed by sonication and centrifuged, applied to NiCl2 Sepahrose resin and eluted with buffer and 500 mM imidazole, pooled, concentrated by ultra centrifugation (10 kDa), buffer exchanged with 50 mM Tris-HCl, pH 8.0, with a PD10 column
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dye-ligand affinity chromatography
-
glutathione Sepharose 4B column chromatography
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Hi-Trap chelating metal affinity column chromatography
leaves of 8 week old field grown plants are ground in liquid nitrogen and extracted with 25 mM NaP buffer, pH 8.0, containing 1 mM EDTA, 5 mM DTT, and 0.5 mM phenylmethanesulfonylfluoride, centrifuged, supernatant taken for enzyme assay
-
Ni2+-NTA resin chromatography
packed red blood cells are washed in phosphate buffered saline, lysed with buffer containing 0.1 M KHEPES, pH 7.5, 0.1% Triton X-100, 1 mM DTT, and 0.02% sodium azide, liver and spleen are diced and homogenized in buffer containing 50 mM KHEPES, pH 7.5, 150 mM KCl, 1 mM Na2EDTA, 1 mM DTT, 0.1 mM PMSF, for activity and thermostability experiments enzymes are expressed in Escherichia coli, cells are harvested and resuspended in 50 mM HEPES, pH 7.8, 0.5 M NaCl, 5 mM DTT, 330 microg/ml lysozyme, 5 microg/ml DNAse and RNAse, freeze-thawed 3times in dry-ice/ethanol, centrifuged, adjusted to pH 7.4 and 8.0 with NaOH, incubated at 37°C or 50°C for 1 h; packed red blood cells are washed in phosphate buffered saline, lysed with buffer containing 0.1 M KHEPES, pH 7.5, 0.1% Triton X-100, 1 mM DTT, and 0.02% sodium azide, liver and spleen are diced and homogenized in buffer containing 50 mM KHEPES, pH 7.5, 150 mM KCl, 1 mM Na2EDTA, 1 mM DTT, 0.1 mM PMSF, for activity and thermostability experiments enzymes are expressed in Escherichia coli, cells are harvested and resuspended in 50 mM HEPES, pH 7.8, 0.5 M NaCl, 5 mM DTT, 330 microg/ml lysozyme, 5 microg/ml DNAse and RNAse, freeze-thawed 3times in dry-ice/ethanol, centrifuged, adjusted to pH 7.4 and 8.0 with NaOH, incubated at 37°C or 50°C for 1 h; packed red blood cells are washed in phosphate buffered saline, lysed with buffer containing 0.1 M KHEPES, pH 7.5, 0.1% Triton X-100, 1 mM DTT, and 0.02% sodium azide, liver and spleen are diced and homogenized in buffer containing 50 mM KHEPES, pH 7.5, 150 mM KCl, 1 mM Na2EDTA, 1 mM DTT, 0.1 mM PMSF, for activity and thermostability experiments enzymes are expressed in Escherichia coli, cells are harvested and resuspended in 50 mM HEPES, pH 7.8, 0.5 M NaCl, 5 mM DTT, 330 microg/ml lysozyme, 5 microg/ml DNAse and RNAse, freeze-thawed 3times in dry-ice/ethanol, centrifuged, adjusted to pH 7.4 and 8.0 with NaOH, incubated at 37°C or 50°C for 1 h
partial
recombinant enzyme from Escherichia coli by ion exchange chromatography and gel filtration
recombinant GST-tagged isoallelic forms K210 and E210 mutants from Escherichia coli to homogeneity
-
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli BL21 (DE3) by glutathione affinity chromatography, the GST-tag is cleaved off
-
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) pLysS by glutathione affinity chromatgraphy, tag cleavage by thrombin, and ultrafiltration
-
recombinant His-tagged enzyme from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography, removal of the His-tag by thrombin and tag elimination by another step of nickel affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli, removal of the His-tag
-
SeMet-labelled enzyme, i.e. [SeMet] HMBS, and wild-type enzyme
-
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and sequencing of naturally occurring mutants G11R and R173Q
-
Escherichia coli BL21(DE3) pLysS with pUHD2 plasmid
-
expressed in Escherichia coli
expressed in Escherichia coli BL21 cells
-
expressed in Escherichia coli strain BL21(DE3)
expression in Escherichia coli BL21(DE3) with pGEX-4T-1 expression vector
-
expression in Escherichia coli BL21+; expression in Escherichia coli BL21+; expression in Escherichia coli BL21+
expression of wild-type and mutant enzymes in SH-SY5Y neuroblastoma cells, expression analysis
-
expression of wild-type and mutants in Escherichia coli strain BL21
-
gene hemC, a single gene for PBGD in chromosome 5, recombinant expression of a codon-optimized PBGD gene in Escherichia coli
gene hemC, recombinant expression of His-tagged enzyme in Escherichia coli
-
gene hemC, recombinant expression of His-tagged enzyme in Escherichia coli strain Rosetta (DE3)
-
Lambda ZAP Express system
-
localization of the porphobilinogen deaminase gene on chromosome 11q23.3
-
PBGD gene, DNA and amino acid sequence determination and analysis, expression of GST-tagged wild-type and mutant enzymes in Escherichia coli BL21 (DE3)
-
PCR-amplification, expression in Escherichia coli BL21Gold(DE3)(pLysS) with plasmid pET-14b(+)hemC
-
pTRE-EalAAT-hPBGD-WPRE plasmid from pTRE2 vector expressed in mice hepatocytes
recombinant expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) pLysS
-
SeMet-labelled enzyme
-
the enzyme is expressed into a housekeeping or an erythroid form as a result of differential promoter usage and splicing, three pairs of isoallelic forms, expression of two of the isoallelic forms, K210 and E210, with mutations involved in acute intermittent porphyria, AIP, in Escherichia coli as GST fusion proteins
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
cf1 mutation show strongly decreased expression compared to wild type
-
transcritpion is upregulated by reactive nitrogen species
transfer of therapeutic plasmid with human enzyme into hepatocytes
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K55Q
-
K55Q mutant
K55Q/K59Q
-
K55Q-K59Q mutant, lower specific activity than the wild-type enzyme
K59Q
-
K59Q mutant, lower specific activity than the wild-type enzyme
A84T
type OregonAR (c250G>A), autosomal recessive, therefore phenotype is homozygous, about 35% wild type activity; type OregonAR (c250G>A), autosomal recessive, therefore phenotype is homozygous, about 35% wild type activity; type OregonAR (c250G>A), autosomal recessive, therefore phenotype is homozygous, about 35% wild type activity
L64S
c.189dupT mutant type Saskatchewan with duplication that leads to a frameshift and thus to a trunkated enzyme due to a premature stop codon 65 and the exchange L64S, autosomal dominant; c.189dupT mutant type Saskatchewan with duplication that leads to a frameshift and thus to a trunkated enzyme due to a premature stop codon 65 and the exchange L64S, autosomal dominant; c.189dupT mutant type Saskatchewan with duplication that leads to a frameshift and thus to a trunkated enzyme due to a premature stop codon 65 and the exchange L64S, autosomal dominant
R149W
type Missouris (c.445C>T), substitution identical to human mutation but there leading to an nonsense mutation, autosomal dominant, less than 1% wild type activity; type Missouris (c.445C>T), substitution identical to human mutation but there leading to an nonsense mutation, autosomal dominant, less than 1% wild type activity; type Missouris (c.445C>T), substitution identical to human mutation but there leading to an nonsense mutation, autosomal dominant, less than 1% wild type activity
887insA
-
site-directed mutagenesis, the mutant shows reduced expression and subcellular distribution compared to the wild-type enzyme
D99G
-
site-directed mutagenesis, inactive holo-enzyme mutant existing as a complex with 2 substrate molecules covalently bound to the dipyrromethane cofactor
G24S
-
c.70G>A (p.Gly24Ser)
G748A
-
site-directed mutagenesis, the mutant shows reduced expression and subcellular distribution compared to the wild-type enzyme
G748C
-
site-directed mutagenesis, the mutant shows reduced expression and subcellular distribution compared to the wild-type enzyme
H300L
-
c.899_900delinsTGCCTGCATCTG (p.His300LeuFsX10)
K132N
-
site-directed mutagenesis, the mutant shows no conformational or kinetic defect, no loss in relative activity (97% of wild-type activity) at standard conditions nor change in Vmax and Km. The mutation is not associated to acute intermittent porphyria, AIP
N322K
-
c.965_966insA (p.Asn322LysfsX36)
Q204K
-
c.610C>A, exon 10, missense mutation, 46% wild type activity, 50 mM Tris-HCl, pH 8.2, 0.1% bovine serum albumin, 0.1% Triton, pH 8.2, 37°C, 1 h in the dark
R116W
-
site-directed mutagenesis, the mutant shows 0.5% of wild-type activity and defects in conformational stability. The mutation is associated to acute intermittent porphyria, AIP
R149Q
-
site-directed mutagenesis, inactive apo-enzyme which is unable to assemble with the dipyrromethane cofactor, the mutant enzyme is unstable and heat-labile
R149X
-
identification of a nonsense mutation in the PBGD gene on chromosome 11q23.3, which harbors the mutations causing acute intermittent porphyria, as the underlying genetic defect in Chester porphyria, phenotype, overview
R167W
-
site-directed mutagenesis, the mutant shows 4.2% of wild-type activity and defects in enzyme kinetics associated with a very high Km and decreased Vmax. The mutation is associated to acute intermittent porphyria, AIP
R173Q/Q204K
-
R173Q is more severe with a resulting enzyme activity of nearly zero, the Q204K increases the negative effect, particularly on the protein stability, 50 mM Tris-HCl, pH 8.2, 0.1% bovine serum albumin, 0.1% Triton, pH 8.2, 37°C, 1 h in the dark
R173W
-
site-directed mutagenesis, the mutant shows 0.6% of wild-type activity and defects in conformational stability and in enzyme kinetics. The mutation is associated to acute intermittent porphyria, AIP
R26H
-
c.77G>A, exon 3, 0.2% wild type activity, 50 mM Tris-HCl, pH 8.2, 0.1% bovine serum albumin, 0.1% Triton, pH 8.2, 37°C, 1 h in the dark
R325A
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c.972_973insG (p.Arg325AlafsX33)
R32P
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c.95G>C (p.Arg32Pro)
R73Q/Q204K
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a complex monoallelic mutation c.[518G>A; c.610C>A] (p.[Arg173Gln;p.Gln204Lys])
V215E
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site-directed mutagenesis, the mutant shows 30% of wild-type activity and lower conformational stability and probably a perturbed elongation process, also 70% loss in both activity and Vmax. The mutation is associated to acute intermittent porphyria, AIP
V215M
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19.4% residual activity compared to the wild type enzyme
L116K
the specific activity of this recombinant mutant enzyme is 5fold higher than that of the recombinant wild type enzyme, catalyzes the formation of uroporphyrinogen I in addition to uroporphyrinogen III
D44A
specific activity is only about 2% of that of the wild type enzyme
E63A
reduced activity
H78L
reduced activity
Q200L
mutant lacks the dipyrromethane cofactor and completely loses the catalytic activity
additional information
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
diagnostics
medicine
pharmacology
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safety, pharmacokinetics and pharmacodynamics of recombinant human porphobilinogen deaminase P 9808, administered to healthy subjects and asymptomatic porphobilinogen deaminase-deficient subjects with high concentrations of porphobilinogen, the substrate of porphobilinogen deaminase for investigation and establishing of an alternative therapy of acute intermittent porphyria, AIP, overview
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