Information on EC 2.3.1.88 - peptide alpha-N-acetyltransferase

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

EC NUMBER
COMMENTARY hide
2.3.1.88
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RECOMMENDED NAME
GeneOntology No.
peptide alpha-N-acetyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
acetyl-CoA + peptide = CoA + Nalpha-acetylpeptide
show the reaction diagram
acetylates N-terminal alanine, serine, methionine and glutamate residues in a number of peptides and proteins, including beta-endorphin, corticotropins and melanotropin. cf. EC 2.3.1.108 tubulin N-acetyltransferase
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetylation
-
-
-
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Acyl group transfer
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:peptide Nalpha-acetyltransferase
Acetylates N-terminal alanine, serine, methionine and glutamate residues in a number of peptides and proteins, including beta-endorphin, corticotropins and melanotropin. cf. EC 2.3.1.108 alpha-tubulin N-acetyltransferase.
CAS REGISTRY NUMBER
COMMENTARY hide
116155-74-9
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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-
-
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Uniprot
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
pig
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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the N-terminal sequences of human H2A and H4 are highly similar to each other and to yeast H4 while the N-terminal sequence of yeast H2A differs, a corresponding mutant cannot be complemented by expression of the human enzyme, overview. Naa40p seems to have co-evolved with the human H2A sequence. Human Naa40p/NatD is conserved from Saccharomyces cerevisiae
malfunction
metabolism
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during eukaryotic translation, the initiator methionine residue is removed from the nascent polypeptide by methionine amino peptidases. The newly exposed N-terminal residue is then modified on its alpha-amino group by transfer of an acetyl group from acetyl-CoA while the polypeptide chain is between 25 to 50 residues long. This modification neutralizes positive a charge on the protein and is thought to influence protein function, stability and interactions with other proteins as well as subsequent modifications
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + AAKKRG
CoA + Nalpha-Ac-AAKKRG
show the reaction diagram
acetyl-CoA + acetyl-beta-endorphin
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + acetyl-CoA synthetase
N-alpha-acetyl-[acetyl-CoA synthetase] + CoA
show the reaction diagram
acetyl-CoA + ADH I-(1-24)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + adrenocorticotropic hormone 1-10
CoA + ?
show the reaction diagram
-
adrenocorticotropic hormone, synthetic peptide
-
-
?
acetyl-CoA + adrenocorticotropic hormone 1-13-NH2
CoA + alphaMSH
show the reaction diagram
acetyl-CoA + adrenocorticotropic hormone 1-18-NH2
CoA + ?
show the reaction diagram
-
corticotropin, synthetic peptide
-
-
?
acetyl-CoA + adrenocorticotropic hormone 1-24
CoA + ?
show the reaction diagram
acetyl-CoA + adrenocorticotropic hormone 1-39
CoA + ?
show the reaction diagram
acetyl-CoA + adrenocorticotropic hormone 1-8
CoA + ?
show the reaction diagram
acetyl-CoA + adrenocorticotropic hormone 4-10
CoA + ?
show the reaction diagram
-
adrenocorticotropic hormone, synthetic peptide
-
-
?
acetyl-CoA + adrenocorticotropic hormone 5-24
CoA + ?
show the reaction diagram
-
corticotropin, synthetic peptide
-
-
?
acetyl-CoA + alpha-endorphin (1-16)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + alpha-melanotropin (1-27)
CoA + alpha-N,O-diacetyl-alpha-melanophore-stimulating hormone
show the reaction diagram
acetyl-CoA + ATPase inhibitor (1-24)(yeast, mitochondrial)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + AVFAD
CoA + Ac-AVFAD
show the reaction diagram
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very low activity with hNaa50p
-
-
?
acetyl-CoA + beta-endorphin (1-27)
CoA + alpha-N-acetyl-beta-endorphin
show the reaction diagram
acetyl-CoA + beta-endorphin (1-31)
CoA + alpha-N-acetyl-beta-endorphin
show the reaction diagram
-
-
-
-
?
acetyl-CoA + camel beta-endorphin (1-31)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + corticotropin(ACTH 1-24)
CoA + alpha-N-acetyl-corticotropin(ACTH 1-24)
show the reaction diagram
-
-
-
?
acetyl-CoA + cytochrome c oxidase (1-24)(yeast, mitochondrial subunit VI)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + DDDIA
CoA + Ac-DDDIA
show the reaction diagram
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beta-actin N-terminus sequence, low activity with hNaa50p
-
-
?
acetyl-CoA + DDDIAAL
CoA + Nalpha-Ac-DDDIAAL
show the reaction diagram
acetyl-CoA + EEEIA
CoA + Ac-EEEIA
show the reaction diagram
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acidic gamma-actin N-terminus sequence, preferred peptide substrate, very low activity with hNaa50p
-
-
?
acetyl-CoA + EEEIAAL
CoA + Nalpha-Ac-EEEIAAL
show the reaction diagram
acetyl-CoA + histone H2A
Nalpha-acetyl-[histone H2A] + CoA
show the reaction diagram
-
-
-
-
?
acetyl-CoA + histone H3
Nalpha-acetyl-[histone H3] + CoA
show the reaction diagram
-
-
-
-
?
acetyl-CoA + human beta-endorphin (1-31)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + human superoxide dismutase (1-24)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + L-alanyl-[peptide]
CoA + N2-acetyl-L-alanyl-[peptide]
show the reaction diagram
acetyl-CoA + L-seryl-[peptide]
CoA + N2-acetyl-L-seryl-[peptide]
show the reaction diagram
acetyl-CoA + L-valyl-[peptide]
CoA + N2-acetyl-L-valyl-[peptide]
show the reaction diagram
acetyl-CoA + MAPLDLD
CoA + Nalpha-Ac-MAPLDLD
show the reaction diagram
acetyl-CoA + MEEKVG
CoA + Nalpha-Ac-MEEKVG
show the reaction diagram
acetyl-CoA + Met-Glu-[peptide]
CoA + Ac-Met-Glu-[peptide]
show the reaction diagram
acetyl-CoA + Met-Leu-[peptide]
CoA + Ac-Met-Leu-[peptide]
show the reaction diagram
-
-
-
?
acetyl-CoA + microtubule
?
show the reaction diagram
-
-
-
?
acetyl-CoA + MLGPE
CoA + Ac-MLGPE
show the reaction diagram
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highly preferred peptide substrate of hNaa50p, very low activity with NatA
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-
?
acetyl-CoA + MLGPEGG
CoA + Nalpha-Ac-MLGPEGG
show the reaction diagram
acetyl-CoA + peptide
CoA + ?
show the reaction diagram
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synthetic oligopeptide representing an acetylated protein as substrate is acetylated in an in vitro HeLa assay by the hNaa16p-hNaa10p-complex, 37°C, 50 mM Tris-HCl, pH 8.5
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-
?
acetyl-CoA + peptide
CoA + Nalpha-acetylpeptide
show the reaction diagram
acetyl-CoA + peptide
Nalpha-acetylpeptide + CoA
show the reaction diagram
-
-
-
?
acetyl-CoA + SESSS
CoA + Ac-SESSS
show the reaction diagram
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N-terminus sequence of an in vivo fully Nalpha-acetylated protein, higher activity peptide substrate of NatA, very low activity with hNaa50p
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-
?
acetyl-CoA + SESSSKS
CoA + Nalpha-Ac-SESSSKS
show the reaction diagram
acetyl-CoA + SSGTPT
CoA + Nalpha-Ac-SSGTPT
show the reaction diagram
acetyl-CoA + viral coat protein of Cabbage leaf curl virus
Nalpha-acetyl-[viral coat protein of Cabbage leaf curl virus ] + CoA
show the reaction diagram
acetyl-CoA + yeast alcohol dehydrogenase I (1-24)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + yeast alcohol dehydrogenase II (1-24)
CoA + ?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + [D-Ser1,Lys17,18]-adrenocorticotropic hormone 1-18-NH2
CoA + ?
show the reaction diagram
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corticotropin, synthetic peptide
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-
?
acetyl-CoA + [Gly1] adrenocorticotropic hormone 1-18-NH2
CoA + ?
show the reaction diagram
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corticotropin, synthetic peptide
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-
?
acetyl-CoA + [N6-PTC-Lys11,15,16,21]-adrenocorticotropic hormone 2-24
CoA + ?
show the reaction diagram
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corticotropin, synthetic peptide
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-
?
acetyl-CoA + [Phe2] ACTH(1-24)
CoA + ?
show the reaction diagram
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-
-
-
?
propionyl-CoA + DDDIAAL
CoA + Nalpha-propionyl-DDDIAAL
show the reaction diagram
propionyl-CoA + EEEIAAL
CoA + Nalpha-propionyl-EEEIAAL
show the reaction diagram
propionyl-CoA + MAPLDLD
CoA + Nalpha-propionyl-MAPLDLD
show the reaction diagram
propionyl-CoA + MLGPEGG
CoA + Nalpha-propionyl-MLGPEGG
show the reaction diagram
propionyl-CoA + SESSSKS
CoA + Nalpha-propionyl-SESSSKS
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
acetyl-CoA + acetyl-CoA synthetase
N-alpha-acetyl-[acetyl-CoA synthetase] + CoA
show the reaction diagram
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acetylation of Lys609. In concert with the CobB sirtuin, the protein acetyltransferase regulates the activity of the central metabolic enzyme acetyl-coenzyme A synthetase
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?
acetyl-CoA + peptide
CoA + Nalpha-acetylpeptide
show the reaction diagram
acetyl-CoA + peptide
Nalpha-acetylpeptide + CoA
show the reaction diagram
Q8R2U5
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-
-
?
acetyl-CoA + viral coat protein of Cabbage leaf curl virus
Nalpha-acetyl-[viral coat protein of Cabbage leaf curl virus ] + CoA
show the reaction diagram
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the acetyltransferase interacts directly with geminivirus movement protein NSP encoded by Cabbage leaf curl virus. The acetyltransferase regulates the nuclear export of the viral genome and potentially other nontranscriptional nuclear events in plant cells
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?
additional information
?
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Q6DBY2
the EGAP NAT complex is an essential regulatory enzyme controlling the function of a subset of proteins required for embryonic growth control and vessel
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetyl-CoA
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetyl-beta-endorphin(1-27)
adrenocorticotropic hormone (11-24)
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inhibits NH2-terminal acetylation of adrenocorticotropic hormone (1-13)NH2
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alpha-N,O-Diacetyl-alphaMSH
diethyldicarbonate
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iodoacetamide
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iodoacetic acid
N-bromosuccinimide
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N-ethylmaleimide
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NH4+
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partially inhibitory
p-chloromercuribenzoate
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Ser-Tyr
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inhibits NH2-terminal acetylation of adrenocorticotropic hormone (1-13)NH2
siRNA
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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cysteine
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dithiothreitol
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glutathione
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.067 - 0.467
acetyl-CoA
0.0022 - 0.008
acetylCoA
0.096 - 0.276
adrenocorticotropic hormone (1-10)
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0.035 - 0.16
adrenocorticotropic hormone (1-13)NH2
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0.024
adrenocorticotropic hormone (1-16)NH2
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0.011
adrenocorticotropic hormone (1-18)
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0.0042 - 0.2
adrenocorticotropic hormone (1-24)
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0.05 - 0.286
adrenocorticotropic hormone (1-39)
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0.037
adrenocorticotropic hormone (4-10)NH2
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0.043
beta-endorphin(1-27)
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0.065
beta-endorphin(1-31)
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.48 - 0.58
acetyl-CoA
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.03 - 8.35
acetyl-CoA
29
0.04 - 0.323
MLGPEGGRWG
169954
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
acetyl-beta-endorphin(1-27)
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0.113
adrenocorticotropic hormone (1-2)[ser-tyr]
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0.106
adrenocorticotropic hormone (1-24)
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0.253
alpha-N,O-Diacetyl-alphaMSH
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000001
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serum
0.0000102
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heart
0.0000142
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kidney
0.0000166
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liver
0.0000169
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lens
0.0000221
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lung
0.000029
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muscle
0.0000378
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brain
0.0000383
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pituitary lobe, posterior-intermediate
0.0000654
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pituitary lobe, anterior
0.000093
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pituitary
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7.5
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7.4 - 8
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2 - 9.8
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
activity assay
37
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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anaplastic thyroid carcinoma
Manually annotated by BRENDA team
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higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
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anaplastic thyroid carcinoma
Manually annotated by BRENDA team
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7-11times more hNAA15 than hNAA16 is expressed, papillary thyroid carcinoma
Manually annotated by BRENDA team
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papillary thyroid carcinoma
Manually annotated by BRENDA team
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anaplastic thyroid carcinoma
Manually annotated by BRENDA team
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follicular thyroid carcinoma
Manually annotated by BRENDA team
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colon carcinoma
Manually annotated by BRENDA team
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5times more hNAA15 than hNAA16 is expressed, high expression, kidney adenocarcinoma
Manually annotated by BRENDA team
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2-3times more hNAA15 than hNAA16 is expressed, high expression, hepatocellular carcinoma, embryonal kidney ATCC CRL-1573
Manually annotated by BRENDA team
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higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
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breast adenocarcinoma
Manually annotated by BRENDA team
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non-small cell lung carcinoma
Manually annotated by BRENDA team
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papillary thyroid carcinoma
Manually annotated by BRENDA team
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primary thyroid cells
Manually annotated by BRENDA team
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papillary thyroid carcinoma
Manually annotated by BRENDA team
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embryonic cell, enzyme is expressed in proliferating cells. Treatment of the cells with retinoic acid initiates exit from the cell cycleand down-regulation of mNAT1 and mARD1
Manually annotated by BRENDA team
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high expression, malignant melanoma
Manually annotated by BRENDA team
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7-11times more hNAA15 than hNAA16 is expressed, neuroepithelioma
Manually annotated by BRENDA team
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2-3times more hNAA15 than hNAA16 is expressed, primary thyroid cells
Manually annotated by BRENDA team
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higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
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higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
additional information
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GaMG cell, 7-11times more hNAA15 than hNAA16 is expressed, glioblastoma; hNAA15 is slightly or significantly overexpressed in all types of thyroid cancer cell lines tested as compared to primary thyroid cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000
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1 * 98000 (Nat1p) + 1 * 27000 (Ard1p) + 1 * 20000 (Nat5p)
27000
-
1 * 98000 (Nat1p) + 1 * 27000 (Ard1p) + 1 * 20000 (Nat5p)
30000
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x * 30000 + x * 100000, ARD1 and NATH form a stable complex, SDs-PAGE
79000
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2 * 79000 + 1 * 83000, 2 protein subunits, 1 RNA subunit, heterotrimer, SDS-PAGE, formamide-PAGE
83000
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2 * 79000 + 1 * 83000, 2 protein subunits, 1 RNA subunit, heterotrimer, SDS-PAGE, formamide-PAGE
98000
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1 * 98000 (Nat1p) + 1 * 27000 (Ard1p) + 1 * 20000 (Nat5p)
98575
-
2 * 98575, cDNA sequence
98800
-
2 * 98800, SDS-PAGE
100000
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x * 30000 + x * 100000, ARD1 and NATH form a stable complex, SDs-PAGE
170000
-
-
180000
-
gel filtration
190000
-
gel filtration
240000
-
gel filtration
241000
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holoenzyme, calculated on the basis of the subunits
250000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 30000 + x * 100000, ARD1 and NATH form a stable complex, SDs-PAGE
trimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetylation
isoform hARD1235 shows autoacetylation
acylation
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the purified protein is crystallized in complex with CoA and with CoA/peptide by sitting-drop vapour-diffusion method at 10°C. The crystals of the 2 complexes belong to the orthorhombic P212121 space group with similar unit cell parameters, and the crystallographic asymmetric unit of both structures contained one molecule only. The crystal structure of the SsArd1-CoA binary complex is determined at 2.13 A resolution and the SsArd1–CoA/peptide ternary complex at 1.84 A
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hanging drop vapour-diffusion method, Ta0058 protein is crystallized at 24°C using a reservoir solution consisting of 0.1 M sodium acetate pH 4.6, 8%(w/v) polyethylene glycol 4000 and 35% (v/v) glycerol. X-ray diffraction data are collected to 2.17 A. The Ta0058 crystals belong to space group P4(1) or P4(3), with unit-cell parameters a = b = 49.3, c = 70.4 A, alpha = beta = gamma = 90°; hanging drop vapour-diffusion method, Ta1140 is crystallized at 24°C using a reservoir solution consisting of 0.1 M trisodium citrate pH 5.6, 20%(v/v) 2-propanol, 20% (w/v) polyethylene glycol 4000 and 0.2 M sodium chloride. X-ray diffraction data are collected to 2.40 A. The Ta1140 crystals belong to space group R3, with hexagonal unit-cell parameters a = b = 75.2, c = 179.6 A, alpha = beta = 90°, gamma = 120°
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to 2.4 A resolution. The structure exhibits the topology beta1-alpha1-beta2-alpha2-beta3-beta4. The N-terminus of Alba interacts with acetyl coenzyme A
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
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about 25% maximal activity at pH 6.0 and 8.0
487649
7.6
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pronounced decline in reaction rate at pH values above
487650
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
-
irreversible denaturation after 1 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme is completely inactivated by lyophilization
-
greatly stablized by inclusion of EDTA and 0.01% deoxycholate in the isolation buffer
-
loses activity by digestion with bovine pancreatic RNase A, Staphylococcus aureus nuclease or proteinase K
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, enzyme is completely inactivated by freezing
-
-20°C, stored as Percoll fraction, activity declines by about 50% in 5 days
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-70°C, 10-20% glycerol, activity is stable for at least 5 months
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-70°C, stored as Percoll fraction, activity declines by about 50% in 5 days
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0-4°C, 10% of the activity is lost after storage for a week
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0°C, sensitive to freezing, more than 90% loss of activity per freeze-thaw cycle
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4°C, stable when stored in 20 mM HEPES buffer, pH 7.4, 0.5 mM dithiothreitol, 10% v,v glycerol, 0.02% NaN3 containing 0.2 M KCl
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4°C, stored as Percoll fraction, activity declines by about 50% in 5 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
extracts from cerebellum are prepared
HEK293 cells are lysed in 50 mM Tris, pH 8.0, containing NaCl, NP-40, EDTA, Na3VO4, and Pefabloc, centrifugation, protein binding unspecifically to agarose beads are removed by centrifugation, immunoprecipitation of hNaa15p and hNaa10p for tryptic LC/MS/MS analysis or SDS-PAGE and Western Blotting. For in vitro enzyme assays transfected HEK293 cell lysate, agarose treated, is incubated with anti-FLAG or unspecific antibody, centrifuged, supernatant incubated with Protein A/G Agarose, centrifuged, and washed with PBS buffer and in acetylation buffer, pH 8.5
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partially
recombinant His-tagged subunits hNaa10p and hNaa50p from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3 N-terminal acetyltransferases, Ard1p/Nat1p3, Nat3p and Mak3p
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cDNA isolation, Xat-1 recombinant protein in vitro translated in rabbit reticulocyte lysate
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cloning and characterization of hNAT5/hSAN, an evolutionarily conserved component of the NatA protein N-alpha-acetyltransferase complex. hNat5 mRNA is expressed in several human cell lines
DNA encoding the wild-type and various mutants of SsArd1 is amplified by PCR and subcloned into a pET28a vector for fusing the gene to a C-terminal His6-tag. The final constructs are verified by DNA sequencing. All of the recombinant plasmids are transformed into Escherichia coli strain BL21 (DE3)
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expression in Escherichia coli
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expression of His-tagged subunits hNaa10p and hNaa50p from vector pETM-41 in Escherichia coli strain BL21(DE3)
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full-length cDNA clone
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gene vnc, molecular mapping and characterization of the wild-type vnc gene and mutant vncBDk gene
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N-terminal acteyltransferases, NatA, NatB and NatC encoded by orthologous genes
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Ta0058 protein is expressed in Escherichia coli Rosetta II(DE3)pLysS cells with an N-terminal purification tag; Ta1140 protein is expressed in Escherichia coli Rosetta II(DE3)pLysS cells with an N-terminal purification tag
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to generate a ARD1-FLAG or a NAT1-Myc expression vector, cDNA is introduced into pcDNA3
transfection of HeLa cells with hNAA16-FLAG and hNAA10-V5 plasmids, and siRNA transfection to produce knockdown of target genes
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E35A
-
activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein) is less than 10% compared to wild-type activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein). 8fold increase in activity with MEEKVG (the N-terminal 6-mer peptide derived from single-stranded DNA-binding protein)
E35F
-
activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein) is less than 10% compared to wild-type activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein). AAKKRG is the N-terminal 6-mer peptide derived from Hjc protein
E35V
-
activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein) is less than 10% compared to wild-type activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein). 5fold increase in activity with MEEKVG (the N-terminal 6-mer peptide derived from single-stranded DNA-binding protein)AAKKRG is the N-terminal 6-mer peptide derived from Hjc protein
E35W
-
activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein) is less than 10% compared to wild-type activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein)
N132A
-
compared to wild-type enzyme 4.5fold increase in Km-value, no significant increase in kcat. The mutant enzyme shows a slight loss in binding affinity for acetyl-CoA with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein)
R100A
-
compared to wild-type enzyme 7fold increase in Km-value, no significant increase in kcat with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein). The R100A mutant shows reduced binding affinity correlated with loss of hydrogen bonds between SsArd1 and AcCoA
T105A
-
compared to wild-type enzyme 3fold increase in Km-value, no significant increase in kcat with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein)
E35A
-
activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein) is less than 10% compared to wild-type activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein). 8fold increase in activity with MEEKVG (the N-terminal 6-mer peptide derived from single-stranded DNA-binding protein)
-
E35V
-
activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein) is less than 10% compared to wild-type activity with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein). 5fold increase in activity with MEEKVG (the N-terminal 6-mer peptide derived from single-stranded DNA-binding protein)AAKKRG is the N-terminal 6-mer peptide derived from Hjc protein
-
N132A
-
compared to wild-type enzyme 4.5fold increase in Km-value, no significant increase in kcat. The mutant enzyme shows a slight loss in binding affinity for acetyl-CoA with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein)
-
R100A
-
compared to wild-type enzyme 7fold increase in Km-value, no significant increase in kcat with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein). The R100A mutant shows reduced binding affinity correlated with loss of hydrogen bonds between SsArd1 and AcCoA
-
T105A
-
compared to wild-type enzyme 3fold increase in Km-value, no significant increase in kcat with SSGTPT (the N-terminal 6-mer peptide derived from Alba protein)
-
additional information
Show AA Sequence (123 entries)
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