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Information on EC 1.14.17.3 - peptidylglycine monooxygenase and Organism(s) Homo sapiens and UniProt Accession P19021
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1.14.17.3 peptidylglycine monooxygenaseIUBMB Comments
A copper protein. The enzyme binds two copper ions with distinct roles during catalysis. Peptidylglycines with a neutral amino acid residue in the penultimate position are the best substrates for the enzyme. The product is unstable and dismutates to glyoxylate and the corresponding desglycine peptide amide, a reaction catalysed by EC 4.3.2.5 peptidylamidoglycolate lyase. In mammals, the two activities are part of a bifunctional protein. Involved in the final step of biosynthesis of alpha-melanotropin and related biologically active peptides.
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Word Map
- 1.14.17.3
-
glycine-extended
- neuropeptides
-
beta-monooxygenase
-
peptidyl-alpha-hydroxyglycine
-
prohormone
- alpha-hydroxyglycine
- peptidylamidoglycolate
-
endoproteolytic
-
neurointermediate
-
corticotrope
-
ascorbate-dependent
-
exafs
-
4.3.2.5
-
4-phenyl-3-butenoic
-
noncoupled
-
side-on
- dbetam
- medicine
- analysis
- synthesis
- pharmacology
The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Synonyms
peptidylglycine alpha-hydroxylating monooxygenase, bifunctional pam, peptidylglycine monooxygenase, peptidyl-glycine alpha-amidating monooxygenase, alpha-ae, peptidylglycine alpha-amidating mono-oxygenase, phmcc, pam-b, peptidylglycine alpha-monooxygenase, peptidylglycine-alpha-amidating monooxygenase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
PAM
PAM-A
-
-
-
-
PAM-B
-
-
-
-
peptide alpha-amidating enzyme
-
-
-
-
peptide alpha-amide synthase
-
-
-
-
peptide-alpha-amide synthetase
-
-
-
-
peptidyl alpha-amidating enzyme
-
-
-
-
peptidylglycine 2-hydroxylase
-
-
-
-
peptidylglycine alpha-amidating monooxygenase
peptidylglycine alpha-hydroxylase
-
-
-
-
peptidylglycine alpha-hydroxylating monooxygenase
synthase, peptide alpha-amide
-
-
-
-
PAM
-
-
-
-
PAM
-
PAM consists of O2-sensitive PHM and peptidyl-alpha-hydroxyglycine alpha-amidating lyase, PAL, activities
peptidylglycine alpha-amidating monooxygenase
-
-
-
-
peptidylglycine alpha-hydroxylating monooxygenase
-
subunit of bifunctional peptidylglycine alpha-amidating monooxygenase (PAM)
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
[peptide]-glycine + 2 ascorbate + O2 = [peptide]-(2S)-2-hydroxyglycine + 2 monodehydroascorbate + H2O
bifunctional enzyme showing peptidylglycine alpha-hydroxylating monooxygenase, EC 1.14.17.3, and peptidylamidoglycolate lyase, PAL, EC 4.3.2.5, activities, the enzyme possesses 2 catalytic domains
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
oxidation
reduction
oxidation
-
-
-
-
reduction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
[peptide]-glycine,ascorbate:oxygen oxidoreductase (2-hydroxylating)
A copper protein. The enzyme binds two copper ions with distinct roles during catalysis. Peptidylglycines with a neutral amino acid residue in the penultimate position are the best substrates for the enzyme. The product is unstable and dismutates to glyoxylate and the corresponding desglycine peptide amide, a reaction catalysed by EC 4.3.2.5 peptidylamidoglycolate lyase. In mammals, the two activities are part of a bifunctional protein. Involved in the final step of biosynthesis of alpha-melanotropin and related biologically active peptides.
CAS REGISTRY NUMBER
COMMENTARY
90597-47-0
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-iodo-Tyr-Val-Gly + ascorbate + O2
D-iodo-Tyr-Val-(2-hydroxyglycine) + dehydroascorbate + H2O
labeled substrate
the carbinol is the substrate for the peptidylamidoglycolate lyase reaction, EC 4.3.2.5, forming glyoxylate and amidated peptide
-
?
hippuric acid + ascorbate + O2
benzamide + glyoxylate + dehydroascorbate + H2O
-
-
-
?
N-dansyl-Tyr-Val-Gly + ascorbate + O2
N-dansyl-Tyr-Val-(2-hydroxyglycine) + dehydroascorbate + H2O
-
-
-
?
N-trinitrophenyl-D-Tyr-Val-Gly + ascorbate + O2
N-trinitrophenyl-D-Tyr-Val-(2-hydroxyglycine) + dehydroascorbate + H2O
-
the carbinol is the substrate for the peptidylamidoglycolate lyase reaction, EC 4.3.2.5, forming glyoxylate and amidated peptide
-
?
proopiomelanocortin peptide + ascorbate + O2
proopiomelanocortin peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
alanyl-L-phenylalanylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
alanylglycyl-L-phenylalanylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
alanylglycyl-L-prolylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
alanylglycylvalyl-L-phenylalanylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
alanylglycylvalyl-L-prolylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
alpha-N-acetyl-Tyr-Val-Gly + ascorbate + O2
alpha-N-acetyl-Tyr-Val-2-hydroxyglycine + dehydroascorbate + H2O
-
-
-
-
?
N-(4-amino-6-methyl-3-oxoheptanoyl)-L-phenylalanylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
N-(4-amino-6-methyl-3-oxoheptanoyl)glycylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
N-acetyl-L-phenylalanylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
N-trinitrophenyl-D-Tyr-Val-Gly + ascorbate + O2
N-trinitrophenyl-D-Tyr-Val-(2-hydroxyglycine) + dehydroascorbate + H2O
-
-
the carbinol is the substrate for the peptidylamidoglycolate lyase reaction, EC 4.3.2.5, forming glyoxylate and amidated peptide
-
?
N-[3-(1-leucylpyrrolidin-2-yl)-3-oxopropanoyl]-L-phenylalanylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
N-[3-(1-leucylpyrrolidin-2-yl)-3-oxopropanoyl]glycylglycine + ascorbate + O2
? + dehydroascorbate + H2O
-
-
-
-
?
peptidylglycine + ascorbate + O2
peptidyl alpha-hydroxyglycine + semidehydroascorbate + H2O
-
-
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
peptidylglycine alpha-hydroxylating monooxygenase reaction
the carbinol is the substrate for the peptidylamidoglycolate lyase reaction, EC 4.3.2.5, forming glyoxylate and amidated peptide
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
most mammalian bioactive peptides possess a C-terminal amino acid amide moiety. Amidated peptides are produced in vivo by the enzymatic cleavage of a precursor with a C-terminal glycine residue. The enzyme catalyzes the key step in the oxidation of the glycine-extended precursors to the alpha-amidated peptide
-
-
?
proopiomelanocortin peptide + ascorbate + O2
proopiomelanocortin peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
a POMC 18-kDa fragment
-
-
?
proopiomelanocortin peptide + ascorbate + O2
proopiomelanocortin peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
a POMC 18-kDa fragment, establishment of an assay method
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
-
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
COOH-terminal glycine
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
amidation of neurohormonal peptides
-
?
additional information
?
-
C-terminally amidation of a range of peptides by the copper-dependent enzyme, peptidylglycine alpha-amidating monooxygenase, PAM
-
-
?
additional information
?
-
constitutive-like secretion of POMC products in recombinant enzyme-expressing cells
-
-
?
additional information
?
-
-
enzyme is required for amidation of autocrine growth factors causing proliferation in tumor cells
-
-
?
additional information
?
-
-
PHM replaces the pro-(S) hydrogen of the carboxy-terminal glycine with a hydroxyl group to form an alpha-hydroxyglycine intermediate
-
-
?
additional information
?
-
-
the enzyme peptidylglycine alpha-amidating monooxygenase consists of two subunits: peptidylglycine alpha-hydroxylating monooxygenase that initiates formation of a radical at a substrate's C-terminal glycine alpha-carbon to then give an alpha-hydroxylated glycine derivative, and peptidylamidoglycolate lyase which catalyzes cleavage of this hydroxyglycine intermediate to give the amidated product with consequent release of glyoxylate
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
proopiomelanocortin peptide + ascorbate + O2
proopiomelanocortin peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
a POMC 18-kDa fragment
-
-
?
peptidylglycine + ascorbate + O2
peptidyl alpha-hydroxyglycine + semidehydroascorbate + H2O
-
-
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
most mammalian bioactive peptides possess a C-terminal amino acid amide moiety. Amidated peptides are produced in vivo by the enzymatic cleavage of a precursor with a C-terminal glycine residue. The enzyme catalyzes the key step in the oxidation of the glycine-extended precursors to the alpha-amidated peptide
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
-
-
-
?
peptidylglycine + ascorbate + O2
peptidyl(2-hydroxyglycine) + dehydroascorbate + H2O
-
amidation of neurohormonal peptides
-
?
additional information
?
-
C-terminally amidation of a range of peptides by the copper-dependent enzyme, peptidylglycine alpha-amidating monooxygenase, PAM
-
-
?
additional information
?
-
constitutive-like secretion of POMC products in recombinant enzyme-expressing cells
-
-
?
additional information
?
-
-
enzyme is required for amidation of autocrine growth factors causing proliferation in tumor cells
-
-
?
additional information
?
-
-
PHM replaces the pro-(S) hydrogen of the carboxy-terminal glycine with a hydroxyl group to form an alpha-hydroxyglycine intermediate
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Cu2+
Zn2+
required for peptidylamidoglycolate lyase activity, 0.62 mol per mol of bifunctional enzyme
copper
Cu2+
Cu2+
required for peptidylglycine alpha-hydroxylating monooxygenase activity, 1.25 mol per mol of bifunctional enzyme
Cu2+
-
The enzyme contains two copper centers, one performs the substrate hydroxylation, while the second is used for electron storage/transfer
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Cinnamic acid
extensive dialysis of bifunctional PAM incubated with cinnamate yielded inactive enzyme unable to catalyze the production of glyoxylate from hippuric acid
([N-[3-(1-leucylpyrrolidin-2-yl)-3-oxopropanoyl]-L-phenylalanyl]oxy)acetic acid
-
-
([[(2S)-1-[[(2-aminopropanoyl)amino]acetyl]pyrrolidin-2-yl]carbonyl]oxy)acetic acid
-
-
4-Phenyl-3-butenoic acid
-
irreversible turnover-dependent inhibition of the enzyme in vitro, at 0.1 mg/ml inhibition in vivo of proliferation of lung cancer cells and Ras-transformed WB rat liver epithelial cells, inhibitor leads to increased gap junction communication in the WB-Ras cells, overview
4-phenyl-3-butenoic acid methyl ester
-
at 0.01 mg/ml inhibition in vivo of proliferation of Ras-transformed WB rat liver epithelial cells, inhibitor leads to increased gap junction communication in the WB-Ras cells, overview
[[(2S)-2-([[(2-aminopropanoyl)amino]acetyl]amino)-3-phenylpropanoyl]oxy]acetic acid
-
-
additional information
cinnamic acid and cinnamic acid analogues are inhibitors or inactivators of PHM. The inactivation chemistry of the cinnamates exhibits all the attributes of a suicide-substrate. But no formation of an irreversible linkage between cinnamate and PHM in the inactivated enzyme is detected. Instead reversible formation of a Michael adduct between an active site nucleophile and cinnamate occurs that leads to inactive enzyme. Cinnamates are found in fruits, fruit juices, vegetables and flowers. Protection of PHM against the cinnamate-mediated inactivation by tiopronin. Molecular docking studies, overview
-
additional information
peptide substrate amidation is strikingly sensitive to the exposure of cells to moderate hypoxia
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.2
alanyl-L-phenylalanylglycine
-
pH and temperature not specified in the publication
0.035
alanylglycyl-L-phenylalanylglycine
-
pH and temperature not specified in the publication
3
alanylglycyl-L-prolylglycine
-
pH and temperature not specified in the publication
0.018
alanylglycylvalyl-L-phenylalanylglycine
-
pH and temperature not specified in the publication
0.96
alanylglycylvalyl-L-prolylglycine
-
pH and temperature not specified in the publication
0.011
N-(4-amino-6-methyl-3-oxoheptanoyl)-L-phenylalanylglycine
-
pH and temperature not specified in the publication
0.86
N-(4-amino-6-methyl-3-oxoheptanoyl)glycylglycine
-
pH and temperature not specified in the publication
0.33
N-(Phenylacetyl)glycine
-
pH and temperature not specified in the publication
0.007
N-[3-(1-leucylpyrrolidin-2-yl)-3-oxopropanoyl]-L-phenylalanylglycine
-
pH and temperature not specified in the publication
0.55
N-[3-(1-leucylpyrrolidin-2-yl)-3-oxopropanoyl]glycylglycine
-
pH and temperature not specified in the publication
0.0033
dansyl-L-Tyr-L-Val-Gly
-
mutant enzyme H107A, in100 mM MES pH 5.5, at 37°C
0.0118
dansyl-L-Tyr-L-Val-Gly
-
mutant enzyme M109I, in100 mM MES pH 5.5, at 37°C
0.0194
dansyl-L-Tyr-L-Val-Gly
-
mutant enzyme H108A, in100 mM MES pH 5.5, at 37°C
0.015
N-acetyl-L-phenylalanylglycine
-
pH and temperature not specified in the publication
0.016
N-acetyl-L-phenylalanylglycine
-
pH and temperature not specified in the publication
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.1
(acetyloxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.007
(decanoylsulfanyl)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.00006
(dodecanoyloxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.37
([(2S)-2-[(2-aminopropanoyl)amino]-3-phenylpropanoyl]oxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.038
([N-[3-(1-leucylpyrrolidin-2-yl)-3-oxopropanoyl]-L-phenylalanyl]oxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.18
([N-[3-(1-leucylpyrrolidin-2-yl)-3-oxopropanoyl]glycyl]oxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.29
([[(2S)-1-(2-aminopropanoyl)pyrrolidin-2-yl]carbonyl]oxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
1.1
([[(2S)-1-[[(2-aminopropanoyl)amino]acetyl]pyrrolidin-2-yl]carbonyl]oxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.2
([[(4-amino-6-methyl-3-oxoheptanoyl)amino]acetyl]oxy)acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.00005
decanoyloxyacetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.002
[(phenylacetyl)oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.045
[(phenylacetyl)sulfanyl]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.045
[[(2S)-2-(acetylamino)-3-phenylpropanoyl]sulfanyl]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.16
[[(2S)-2-([[(2-aminopropanoyl)amino]acetyl]amino)-3-phenylpropanoyl]oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
1.6
[[(2S)-2-amino-3-phenylpropanoyl]oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.5
[[(2S)-pyrrolidin-2-ylcarbonyl]oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
1.8
[[(acetylamino)acetyl]oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.009
[[N-(4-amino-6-methyl-3-oxoheptanoyl)-L-phenylalanyl]oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.006
[[(2S)-2-(acetylamino)-3-phenylpropanoyl]oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.011
[[(2S)-2-(acetylamino)-3-phenylpropanoyl]oxy]acetic acid
Homo sapiens
-
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.53
purified recombinant enzyme, peptidylglycine alpha-hydroxylating monooxygenase activity
additional information
-
peptidylglycine alpha-amidating and alpha-hydroxylating monooxygenase activities in several tumor tissue extracts, comparison with immunological analysis, overview, highest activity in medullary thyroid carcinoma
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.8
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
bifunctional enzyme showing peptidylglycine alpha-hydroxylating monooxygenase, PHM, EC 1.14.17.3, and peptidylamidoglycolate lyase, PAL, EC 4.3.2.5, activities
SwissProt
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
quantification of PAM endocytic trafficking to the trans-Golgi network
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
physiological function
malfunction
disruption of AP-1-dependent late endosomal trafficking diminishes the ability of PAM to retain copper and produce amidated peptides. Impaired AP-1 function alters luminal copper delivery to PAM. Altered luminal cuproenzyme function may contribute to diseases associated with diminished AP-1 function. Reduced AP-1 function makes 18-kDa fragment amidation more sensitive to inhibition by bathocuproine disulfonate, a cell-impermeant Cu(I) chelator. The endocytic trafficking of PAM is altered, and PAM-1 accumulates on the cell surface when AP-1 levels are reduced
malfunction
the rs13175330 polymorphism of the PAM gene is selected from the ten single nucleotide polymorphisms (SNPs) most strongly associated with blood pressure. The presence of the G allele of the PAM rs13175330 A>G SNP is associated with a higher risk of hypertension after adjustments for age, sex, BMI, smoking, and drinking. The PAM rs13175330 A>G SNP is a candidate gene for hypertension in the Korean population. Additionally, the PAM rs13175330 G allele might be associated with insulin resistance and LDL atherogenicity in patients with hypertension. Phenotypes, overview
physiological function
most mammalian bioactive peptides possess a C-terminal amino acid amide moiety. Amidated peptides are produced in vivo by the enzymatic cleavage of a precursor with a C-terminal glycine residue. The enzyme catalyzes the key step in the oxidation of the glycine-extended precursors to the alpha-amidated peptide
physiological function
in neuroendocrine cells, ATP7A provides copper to peptidylglycine alpha-amidating monooxygenase (PAM), an essential enzyme that requires copper to catalyze peptide amidation, one of the final steps in the production of bioactive peptides. Trafficking of Atp7a, a copper pump, and the cuproenzyme PAM-1 depend on adaptor protein-1 complex (AP-1). The enzyme is involved in the prohormone POMC processing pathway and constitutive-like secretion, overview. Production of the amidated products of POMC (18-kDa fragment-NH2, JP-NH2, and ACTH(1-13)NH2) requires both PAM and Atp7a
physiological function
PAM is a bifunctional enzyme, its copper-dependent peptidylglycine alpha-hydroxylating monooxygenase, PHM, domain converts peptidylglycine substrates to peptidyl-alpha-hydroxyglycine intermediates that are subsequently converted into amidated products plus glyoxylate by the zinc-dependent peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) domain. The reaction catalyzed by PHM results in the stereospecific incorporation of one atom of molecular oxygen into the substrate in a reaction that involves two single electron transfer steps. PAM-mediated C-terminal amidation occurs across a range of biologically active endocrine and nervous system peptides and in many cases has been shown to be required for normal biological activity in vivo. Peptidylglycine alpha-amidating monooxygenase (PAM) is solely responsible for catalysis of amidation, a biologically important posttranslational modification. Peptide substrate amidation is strikingly sensitive to the exposure of cells to moderate hypoxia, physiological effects of hypoxia may be PAM-dependent. Because PAM-dependent amidation is irreversible, bi-directional responses that rapidly upregulate and downregulate levels of amidation can only be observed on rapidly turned-over PAM substrates
physiological function
PAM is the only known enzyme responsible for the bioconversion of glycine C-terminal prohormones into des-glycine alpha-amidated products and glyoxylate. PAM is a bifunctional, type-II copper monooxygenase that consists of two independent catalytic domains, PHM and PAL. The PAM reaction is a two-step process. Initially, PHM removes the pro-S hydrogen to allow hydroxylation of the alpha-glycyl carbon, resulting in an alpha-hydroxylated intermediate. PHM is a molecular oxygen, Cu(II) and ascorbate (reductant) dependent enzyme. The second catalytic domain, PAL, dealkylates the alpha-hydroxylated intermediate, yielding the alpha-amidated product and glyoxylate. PAM is responsible for the posttranslational modification of many important neuropeptides, including oxytocin, vasopressin, ACTH, alphaMSH, VIP, substance P, neuropeptide Y, cholecystokinin, gastrin, and a large number of other molecules. Hypoglycemic effect of the alpha-amidated analogue of recombinant human insulin
physiological function
peptidylglycine alpha-amidating monooxygenase (PAM) is a bifunctional enzyme that catalyzes the final reaction in the maturation of alpha-amidated peptide hormones. Peptidylglycine alpha-hydroxylating monooxygenase (PHM) is the PAM domain responsible for the copper-, ascorbate- and O2-dependent hydroxylation of glycine-extended precursor peptides to the active alpha-amidated peptide and glyoxylate. Peptidylamidoglycolate lyase, EC 4.3.2.5, is the PAM domain responsible for the Zn(II)-dependent dealkylation of the alpha-hydroxyglycine-containing precursor to the final alpha-amidated peptide
physiological function
peptidylglycine-alpha-amidating monooxygenase (PAM) may play a role in the secretion of atrial natriuretic peptide (ANP), which is a hormone involved in the maintenance of blood pressure. Enzyme PAM is involved in regulation of blood pressure and hypertension
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). AMD_HUMAN
973
1
108332
Swiss-Prot
Secretory Pathway (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
the specific components of the thioredoxin-hPHMcc-His6 fusion protein are as follows: 11.8 kDa for thioredoxin, 5.9 kDa peptide between thioredoxin and hPHMcc, 35.2 kDa for hPMHcc, a 1.5 kDa peptide on the C-terminal side of hPHMcc, and 0.6 kDa for the His6 tag. The sum of the individual masses are 55 kDa, in agreement with the 52 kDa measure by SDS-PAGE
additional information
-
the specific components of the thioredoxin-hPHMcc-His6 fusion protein are as follows: 11.8 kDa for thioredoxin, 5.9 kDa peptide between thioredoxin and hPHMcc, 35.2 kDa for hPMHcc, a 1.5 kDa peptide on the C-terminal side of hPHMcc, and 0.6 kDa for the His6 tag. The sum of the individual masses are 55 kDa, in agreement with the 52 kDa measure by SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
enzyme contains 2 putative N-glycosylation sites at ASn411 and Asn762, the purified enzyme contains at least 1 sugar chain
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
M314H
-
the catalytic activity of the mutant decreases by 96% due to effects on both kcat and KM but it displayed the same activity/pH profile with a maximum around pH 6.0
additional information
identification of naurally occuring single nucleotide polymorphisms (SNPs), including the rs13175330 polymorphism. The presence of the G allele of the gene PAM rs13175330 A>G SNP is associated with a higher risk of hypertension after adjustments for age, sex, BMI, smoking, and drinking. The rs13175330 G allele carriers in the hypertension group treated without antihypertensive therapy (HTN w/o therapy) have significantly higher systolic and diastolic blood pressure than the AA genotype carriers, whereas the G allele carriers in the hypertension group treated with antihypertensive therapy (HTN w/ therapy) show significantly higher diastolic blood pressure. The rs13175330 G allele carriers in the HTN w/o therapy group have significantly increased levels of insulin, insulin resistance, and oxidized low-density lipoprotein (LDL) and significantly decreased LDL-cholesterol levels and LDL particle sizes compared to the AA carriers
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
oxygen sensitivity of the peptidylglycine alpha-amidating monooxygenase (PAM) in neuroendocrine cells
745332
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant PAM from CHO dhfr cells by ammonium sulfate fractionation and anion exchange chromatography
recombinant soluble enzyme from culture medium in a 3-step chromatographic procedure, about 40fold to near homogeneity
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene PAM, DNA and amino acid sequence determination and analysis, genotyping
gene PAM, stable recombinant expression of PAM in CHO dhfr cells, construction of pCG/dhfr plasmid with added PAMLID gene incorporating the altered leader sequence
production of the catalytic core of human peptidylglycine alpha-hydroxylating monooxygenase (hPHMcc) in Escherichia coli possessing a N-terminal fusion to thioredoxin. Thioredoxin is fused to hPHMcc to enhance the yield of the resulting 52 kDa protein as a soluble and catalytically active enzyme
subcloning from human heart lambda-gt10 library in Escherichia coli, soluble recombinant enzyme through elimination of the C-terminal membrane-spanning domain, expression of a soluble form in CHO K-1 cells, excretion of the recombinant enzyme to the cell culture medium
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
enzyme PAM-dependent amidation has the potential to signal oxygen levels in the same range as the hypoxia-inducible factor (HIF) system
synthesis
usage of the recombinant PAM for insulin analogue amidation producing insulin glargine amide, an insulin derivative that shows a time/effect profile which is distinctly more flat and thus more advantageous than insulin glargine itself. The enzyme is used to modify glycine-extended A22(G)-B31(K)-B32(R) human insulin analogue (GKR). Hypoglycemic activity of amidated and non-amidated insulin is compared
medicine
medicine
-
PHM is a potential target for the development of inhibitors as drugs for the teratment of human disease
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Scopsi, L.; Lee, R.; Gullo, M.; Collini, P.; Husten, E.J.; Eipper, B.A.
Peptidylglycine alpha-amidating monooxygenase in neuroendocrine tumors: its identification, characterization, quantification, and relation to the grade of morphologic differentiation, amidated peptide content, and granin immunocytochemistry
Appl. Immunohistochem.
6
120-132
1998
Homo sapiens
-
Satani, M.; Takahashi, K.; Sakamoto, H.; Harada, S.; Kaida, Y.; Noguchi, M.
Expression and characterization of human bifunctional peptidylglycine alpha-amidating monooxygenase
Protein Expr. Purif.
28
293-302
2003
Homo sapiens (P19021), Homo sapiens
Sunman, J.A.; Foster, M.S.; Folse, S.L.; May, S.W.; Matesic, D.F.
Reversal of the transformed phenotype and inhibition of peptidylglycine alpha-monooxygenase in Ras-transformed cells by 4-phenyl-3-butenoic acid
Mol. Carcinog.
41
231-246
2004
Homo sapiens
Klinman, J.P.
The copper-enzyme family of dopamine b-monooxygenase and peptidylglycine a-hydroxylating monooxygenase: Resolving the chemical pathway for substrate hydroxylation
J. Biol. Chem.
281
3013-3016
2006
Homo sapiens
Merkler, D.J.; Asser, A.S.; Baumgart, L.E.; Carballo, N.; Carpenter, S.E.; Chew, G.H.; Cosner, C.C.; Dusi, J.; Galloway, L.C.; Lowe, A.B.; Lowe, E.W.; King, L.; Kendig, R.D.; Kline, P.C.; Malka, R.; Merkler, K.A.; McIntyre, N.R.; Romero, M.; Wilcox, B.J.; Owen, T.C.
Substituted hippurates and hippurate analogs as substrates and inhibitors of peptidylglycine alpha-hydroxylating monooxygenase (PHM)
Bioorg. Med. Chem.
16
10061-10074
2008
Blattella germanica, Homo sapiens, Rattus norvegicus (P14925)
Trendel, J.A.; Ellis, N.; Sarver, J.G.; Klis, W.A.; Dhananjeyan, M.; Bykowski, C.A.; Reese, M.D.; Erhardt, P.W.
Catalytically active peptidylglycine alpha-amidating monooxygenase in the media of androgen-independent prostate cancer cell lines
J. Biomol. Screen.
13
804-809
2008
Homo sapiens
Handa, S.; Spradling, T.J.; Dempsey, D.R.; Merkler, D.J.
Production of the catalytic core of human peptidylglycine alpha-hydroxylating monooxygenase (hPHMcc) in Escherichia coli
Protein Expr. Purif.
84
9-13
2012
Homo sapiens (P19021), Homo sapiens
Bauman, A.T.; Broers, B.A.; Kline, C.D.; Blackburn, N.J.
A copper-methionine interaction controls the pH-dependent activation of peptidylglycine monooxygenase
Biochemistry
50
10819-10828
2011
Homo sapiens
Kline, C.D.; Mayfield, M.; Blackburn, N.J.
HHM motif at the CuH-site of peptidylglycine monooxygenase is a pH-dependent conformational switch
Biochemistry
52
2586-2596
2013
Homo sapiens
Morris, K.M.; Cao, F.; Onagi, H.; Altamore, T.M.; Gamble, A.B.; Easton, C.J.
Prohormone-substrate peptide sequence recognition by peptidylglycine alpha-amidating monooxygenase and its reflection in increased glycolate inhibitor potency
Bioorg. Med. Chem. Lett.
22
7015-7018
2012
Homo sapiens
Cao, F.; Gamble, A.; Kim, H.; Onagi, H.; Gresser, M.; Kerr, J.; Easton, C.
Potent and selective inhibitors of human peptidylglycine alpha-amidating monooxygenase
MedChemComm
2
760-763
2011
Homo sapiens
-
Yoo, H.J.; Kim, M.; Kim, M.; Chae, J.S.; Lee, S.H.; Lee, J.H.
The peptidylglycine-alpha-amidating monooxygenase (PAM) gene rs13175330 A>G polymorphism is associated with hypertension in a Korean population
Hum. Genomics
11
29
2017
Homo sapiens (P19021)
Bonnemaison, M.L.; Baeck, N.; Duffy, M.E.; Ralle, M.; Mains, R.E.; Eipper, B.A.
Adaptor protein-1 complex affects the endocytic trafficking and function of peptidylglycine alpha-amidating monooxygenase, a luminal cuproenzyme
J. Biol. Chem.
290
21264-21279
2015
Rattus norvegicus (P14925), Homo sapiens (P19021)
Simpson, P.D.; Eipper, B.A.; Katz, M.J.; Gandara, L.; Wappner, P.; Fischer, R.; Hodson, E.J.; Ratcliffe, P.J.; Masson, N.
Striking oxygen sensitivity of the peptidylglycine alpha-amidating monooxygenase (PAM) in neuroendocrine cells
J. Biol. Chem.
290
24891-24901
2015
Drosophila melanogaster (O01404), Homo sapiens (P19021), Mus musculus (P97467)
McIntyre, N.R.; Lowe, E.W.; Battistini, M.R.; Leahy, J.W.; Merkler, D.J.
Inactivation of peptidylglycine alpha-hydroxylating monooxygenase by cinnamic acid analogs
J. Enzyme Inhib. Med. Chem.
31
551-562
2016
Rattus norvegicus (P14925), Homo sapiens (P19021)
Zielinski, M.; Wojtowicz-Krawiec, A.; Mikiewicz, D.; Kesik-Brodacka, M.; Cecuda-Adamczewska, V.; Marciniak-Rusek, A.; Sokolowska, I.; Lukasiewicz, N.; Gurba, L.; Odrowaz-Sypniewski, M.; Baran, P.; Plucienniczak, G.; Plucienniczak, A.; Borowicz, P.; Szewczyk, B.
Expression of recombinant human bifunctional peptidylglycine alpha-amidating monooxygenase in CHO cells and its use for insulin analogue modification
Protein Expr. Purif.
119
102-109
2016
Homo sapiens (P19021)
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