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Information on EC 4.3.2.5 - peptidylamidoglycolate lyase and Organism(s) Rattus norvegicus and UniProt Accession P14925

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EC Tree
     4 Lyases
         4.3 Carbon-nitrogen lyases
             4.3.2 Amidine-lyases
                4.3.2.5 peptidylamidoglycolate lyase
IUBMB Comments
Requires zinc. The enzyme acts on the product of the reaction catalysed by EC 1.14.17.3 peptidylglycine monooxygenase, thus removing a terminal glycine residue and leaving a des-glycine peptide amide. In mammals, the two activities are part of a bifunctional protein.
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This record set is specific for:
Rattus norvegicus
UNIPROT: P14925
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The taxonomic range for the selected organisms is: Rattus norvegicus
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Synonyms
peptidylamidoglycolate lyase, bifunctional peptidylglycine alpha-amidating enzyme, alpha-hydroxyglycine amidating dealkylase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
bifunctional peptidylglycine alpha-amidating enzyme
-
peptidyl-alpha-hydroxyglycine alpha-amidating lyase
-
rPAL gene product
-
alpha-hydroxyglycine amidating dealkylase
-
-
-
-
lyase, peptidyl-alpha hydroxyglycine
-
-
-
-
peptidyl-alpha-hydroxyglycine alpha-amidating lyase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
amidine-lyase reaction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
[peptide]-(2S)-2-hydroxyglycine peptidyl-amide-lyase (glyoxylate-forming)
Requires zinc. The enzyme acts on the product of the reaction catalysed by EC 1.14.17.3 peptidylglycine monooxygenase, thus removing a terminal glycine residue and leaving a des-glycine peptide amide. In mammals, the two activities are part of a bifunctional protein.
CAS REGISTRY NUMBER
COMMENTARY hide
131689-50-4
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-N-dansyl-Tyr-Val-alpha-hydroxyglycine
N-dansyl-Tyr-Val-NH2 + glyoxylate
show the reaction diagram
-
-
-
?
alpha-N-acetyl-Tyr-Val-alpha-hydroxyglycine
alpha-N-acetyl-Tyr-Val-NH2 + glyoxylate
show the reaction diagram
-
-
-
?
peptidylamidoglycolate
peptidyl amide + glyoxylate
show the reaction diagram
-
-
-
?
alpha-hydroxyglycine-extended peptide
peptidyl amide + glyoxylate
show the reaction diagram
-
-
-
-
?
dansyl-Tyr-Val-alpha-hydroxyglycine
dansyl-Tyr-Val-NH2 + glyoxylate
show the reaction diagram
-
-
-
?
dansyl-Tyr-Val-Gly
dansyl-Tyr-Val-NH2 + glyoxylate
show the reaction diagram
-
-
-
-
?
Nalpha-acetyl-Tyr-Val-alpha-hydroxyglycine
Nalpha-acetyl-Tyr-Val-NH2 + glyoxylate
show the reaction diagram
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
peptidylamidoglycolate
peptidyl amide + glyoxylate
show the reaction diagram
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
no cofactor requirement
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
activates the enzyme
Co2+
activates the enzyme
Zn2+
tightly blound
Zn2+
-
0.7 mol per mol enzyme
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
-
EDTA
-
addition of 0.5-1 M reduces activity to 50%, Ca2+, Mn2+, Co2+ or Ni2+ restore activity
EGTA
-
addition of 0.5-1 M reduces activity to 50%, Ca2+, Mn2+, Co2+ or Ni2+ restore activity
HCl
-
0.5 M guanidinium HCl reduces activity to 28%
o-phenanthroline
-
-
Urea
-
addition of 1 M urea reduces activity to 58%
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.013
alpha-N-Acetyl-Tyr-Val-alpha-hydroxyglycine
-
0.035 - 0.048
Dansyl-Tyr-Val-alpha-hydroxyglycine
-
37°C, pH 7.0
0.0023 - 0.004
dansyl-Tyr-Val-Gly
-
37°C, pH 6
0.0102 - 0.33
Nalpha-Acetyl-Tyr-Val-alpha-hydroxyglycine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
86 - 380
Nalpha-Acetyl-Tyr-Val-alpha-hydroxyglycine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.25 - 0.32
-
dansyl-Tyr-Val-Gly as substrate
4.5 - 8.3
-
dansyl-Tyr-Val-alpha-hydroxyglycine as substrate
57.24
-
affinity chromatography step II
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
enzyme is secreted into the medium from mouse C127 cells transfected with the rat MTC cDNA encoding the truncated type A enzyme
Manually annotated by BRENDA team
-
in peripheral islet and B-cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
AMD_RAT
976
2
108675
Swiss-Prot
Secretory Pathway (Reliability: 4)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38000
-
gel filtration
41000
-
x * 41000, SDS-PAGE
53000
-
gel filtration
75000
-
truncated type A enzyme which catalyzes a two-step reaction involving an initial hydroxylation of peptidyl-Gly followed by conversion of the peptidyl-alpha-hydroxyglycine intermediate to the amidated product
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 41000, SDS-PAGE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D590N
-
no secretion
D599N
-
no secretion
D652N
-
no secretion
D705N
-
much less secretion and less active than wild-type
E707Q
-
much less secretion than wild-type
H532A
-
no secretion
H585A
-
less active than wild-type
H603A
-
no secretion
H690A
-
less active than wild-type
H697A
-
no changes in secretion an activity
H786A
-
less active than wild-type
H817A
-
no changes in secretion an activity
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in a human embryonic kidney cell line that lacks regulated secretory granules
-
expression in Chinese hamster ovary cells
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Merkler, D.J.; Young, S.D.
Recombinant type A rat 75-kDa alpha-amidating enzyme catalyzes the conversion of glycine-extended peptides to peptide amides via an alpha-hydroxyglycine intermediate
Arch. Biochem. Biophys.
289
192-196
1991
Rattus norvegicus
Manually annotated by BRENDA team
Francisco, W.A.; Merkler, D.J.; Blackburn, N.J.; Klinman, J.P.
Kinetic mechanism and intrinsic isotope effects for the peptidylglycine alpha-amidating enzyme reaction
Biochemistry
37
8244-8252
1998
Rattus norvegicus
Manually annotated by BRENDA team
Husten, E.J.; Tausk, F.A.; Keutmann, H.T.; Eipper, B.A.
Use of endoproteases to identify catalytic domains, linker regions, and functional interactions in soluble peptidylglycine alpha-amidating monooxygenase
J. Biol. Chem.
268
9709-9717
1993
Bos taurus, Rattus norvegicus
Manually annotated by BRENDA team
Husten, E.J.; Eipper, B.A.
The membrane-bound bifunctional peptidylglycine alpha-amidating monooxygenase protein. Exploration of its domain structure through limited proteolysis.
J. Biol. Chem.
266
17004-17010
1991
Rattus norvegicus
Manually annotated by BRENDA team
Takahashi, K.; Okamoto, H.; Seino, H.; Noguchi, M.
Peptidylglycine alpha-amidating reaction: evidence for a two-step mechanism involving a stable intermediate at neutral pH
Biochem. Biophys. Res. Commun.
169
524-530
1990
Bos taurus, Rattus norvegicus
Manually annotated by BRENDA team
Kato, I.; Yonekura, H.; Tajima, M.; Yanagi, M.; Yamamoto, H.; Okamoto, H.
Two enzymes concerned in peptide hormone alpha-amidation are synthesized from a single mRN
Biochem. Biophys. Res. Commun.
172
197-203
1990
Rattus norvegicus
Manually annotated by BRENDA team
Kolhekar, A.S.; Bell, J.; Shiozaki, E.N.; Jin, L.; Keutmann, H.T.; Hand, T.A.; Mains, R.E.; Eipper, B.A.
Essential features of the catalytic core of peptidyl-a-hydroxyglycine alpha-amidating Lyase
Biochemistry
41
12384-12394
2002
Rattus norvegicus
Manually annotated by BRENDA team
Garmendia, O.; Rodriguez, M.P.; Burrell, M.A.; Villaro, A.C.
Immunocytochemical finding of the amidating enzymes in mouse pancreatic A-, B-, and D-cells: a comparison with human and rat
J. Histochem. Cytochem.
50
1401-1415
2002
Homo sapiens, Mus musculus, Rattus norvegicus
Manually annotated by BRENDA team
Han, M.; Park, D.; Vanderzalm, P.J.; Mains, R.E.; Eipper, B.A.; Taghert, P.H.
Drosophila uses two distinct neuropeptide amidating enzymes, dPAL1 and dPAL2
J. Neurochem.
90
129-141
2004
Drosophila melanogaster (Q9V5E1), Drosophila melanogaster (Q9W1L5), Rattus norvegicus (P14925)
Manually annotated by BRENDA team
Bell, J.; Ash, D.E.; Snyder, L.M.; Kulathila, R.; Blackburn, N.J.; Merkler, D.J.
Structural and functional investigations on the role of zinc in bifunctional rat peptidylglycine alpha-amidating enzyme
Biochemistry
36
16239-16246
1997
Rattus norvegicus (P14925)
Manually annotated by BRENDA team