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
-
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
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetylation
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
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
pig
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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
-
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 + ACTH peptide
CoA + ?
show the reaction diagram
-
17 amino acids are identical to the adrenocorticotropin (ACTH) peptide sequence, the ACTH-derived lysines are replaced by arginines to minimize any potential interference by Nalpha-acetylation
-
-
?
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
-
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 + CAP2
?
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
-
beta-actin N-terminus sequence, low activity with hNaa50p
-
-
?
acetyl-CoA + DDDIAAL
CoA + Nalpha-Ac-DDDIAAL
show the reaction diagram
acetyl-CoA + DDDIAALRWGRPVGRRRRPVRVYP
CoA + Nalpha-acetyl-DDDIAALRWGRPVGRRRRPVRVYP
show the reaction diagram
-
-
-
-
acetyl-CoA + EEEIA
CoA + Ac-EEEIA
show the reaction diagram
-
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 + EEEIAALRWGRPVGRRRRPVRVYP
CoA + Nalpha-acetyl-EEEIAALRWGRPVGRRRRPVRVYP
show the reaction diagram
-
-
-
-
acetyl-CoA + GRX3
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + histone H2A
CoA + Nalpha-acetyl-histone H2A
show the reaction diagram
Saccharomyces cerevisiae histone H2A, 2SGGKGGKAGSAAKASQSR19 N-terminal sequence
-
-
?
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 + histone H4
CoA + Nalpha-acetyl-histone H4
show the reaction diagram
Saccharomyces cerevisiae histone H4, 2SGRGKGGKGLGKGGAKR18 N-terminal sequence. hNaa40p catalyzes H4 Nalpha-acetylation and not H4 lysine Nepsilon-acetylation, immunoprecipitated hNaa40p specifically Nt-acetylates H4 in vitro
-
-
?
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
-
highly preferred peptide substrate of hNaa50p, very low activity with NatA
-
-
?
acetyl-CoA + MLGPEGG
CoA + Nalpha-Ac-MLGPEGG
show the reaction diagram
acetyl-CoA + MLGPEGGRWG
CoA + Ac-MLGPEGGRWG
show the reaction diagram
-
-
-
-
?
acetyl-CoA + MLGPEGGRWGRPVGRRRRPVRVYP
CoA + Nalpha-acetyl-MLGPEGGRWGRPVGRRRRPVRVYP
show the reaction diagram
-
-
-
-
acetyl-CoA + NTF2
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + Orc1p
CoA + Nalpha-acetyl-[Orc1p]
show the reaction diagram
acetyl-CoA + PCNP protein
CoA + Nalpha-acetyl-PCNP
show the reaction diagram
-
i.e. PEST proteolytic signal-containing nuclear protein
-
-
?
acetyl-CoA + PDC5
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + peptide
CoA + ?
show the reaction diagram
-
synthetic oligopeptide representing an acetylated protein as substrate is acetylated in an in vitro HeLa assay by the hNaa16p-hNaa10p-complex, 37C, 50 mM Tris-HCl, pH 8.5
-
-
?
acetyl-CoA + peptide
CoA + Nalpha-acetylpeptide
show the reaction diagram
acetyl-CoA + peptide
Nalpha-acetylpeptide + CoA
show the reaction diagram
acetyl-CoA + PIL1
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + POL30
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + RAD23
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + RPS12
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + RUP2
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + SASEAGVRWGRPVGRRRRP
CoA + Nalpha-acetyl-SASEAGVRWGRPVGRRRRP
show the reaction diagram
-
-
-
?
acetyl-CoA + SESSS
CoA + Ac-SESSS
show the reaction diagram
-
N-terminus sequence of an in vivo fully Nalpha-acetylated protein, higher activity peptide substrate of NatA, very low activity with hNaa50p
-
-
?
acetyl-CoA + SESSSKS
CoA + Nalpha-Ac-SESSSKS
show the reaction diagram
acetyl-CoA + SESSSKSRWGRPVGRRRRPVRVYP
CoA + Ac-SESSSKSRWGRPVGRRRRPVRVYP
show the reaction diagram
-
high-mobility-group protein A1 sequence
-
-
?
acetyl-CoA + SESSSKSRWGRPVGRRRRPVRVYP
CoA + Nalpha-acetyl-SESSSKSRWGRPVGRRRRPVRVYP
show the reaction diagram
-
-
-
-
acetyl-CoA + silencing facor Sir3p
CoA + Nalpha-acetyl-[silencing factor Sir3p]
show the reaction diagram
acetyl-CoA + SMI1
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + SNZ2/3
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + SSGTPT
CoA + Nalpha-Ac-SSGTPT
show the reaction diagram
acetyl-CoA + THI20
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + THI22
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + THI4
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + THI6
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + TIF6
?
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 + VMA6
?
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 + ZPS1
?
show the reaction diagram
-
-
-
-
?
acetyl-CoA + [D-Ser1,Lys17,18]-adrenocorticotropic hormone 1-18-NH2
CoA + ?
show the reaction diagram
-
corticotropin, synthetic peptide
-
-
?
acetyl-CoA + [Gly1] adrenocorticotropic hormone 1-18-NH2
CoA + ?
show the reaction diagram
-
corticotropin, synthetic peptide
-
-
?
acetyl-CoA + [N6-PTC-Lys11,15,16,21]-adrenocorticotropic hormone 2-24
CoA + ?
show the reaction diagram
-
corticotropin, synthetic peptide
-
-
?
acetyl-CoA + [Phe2] ACTH(1-24)
CoA + ?
show the reaction diagram
-
-
-
-
?
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
?
-
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
-
acetylation of Lys609. In concert with the CobB sirtuin, the protein acetyltransferase regulates the activity of the central metabolic enzyme acetyl-coenzyme A synthetase
-
-
?
acetyl-CoA + MLGPEGGRWG
CoA + Ac-MLGPEGGRWG
show the reaction diagram
-
-
-
-
?
acetyl-CoA + Orc1p
CoA + Nalpha-acetyl-[Orc1p]
show the reaction diagram
-
Orc1p is the large subunit of the origin recognition complex, ORC. Mechanism for modulating chromaffin function
-
-
?
acetyl-CoA + PCNP protein
CoA + Nalpha-acetyl-PCNP
show the reaction diagram
-
i.e. PEST proteolytic signal-containing nuclear protein
-
-
?
acetyl-CoA + peptide
CoA + Nalpha-acetylpeptide
show the reaction diagram
acetyl-CoA + peptide
Nalpha-acetylpeptide + CoA
show the reaction diagram
acetyl-CoA + silencing facor Sir3p
CoA + Nalpha-acetyl-[silencing factor Sir3p]
show the reaction diagram
-
mechanism for modulating chromaffin function
-
-
?
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
-
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
-
-
?
additional information
?
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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)
-
inhibits NH2-terminal acetylation of adrenocorticotropic hormone (1-13)NH2
-
alpha-N,O-Diacetyl-alphaMSH
CoA
-
product inhibition, competitive versus acetyl-CoA, noncompetitive against peptide substrate well in line with a ternary complex mechanism. The non-competitive inhibition pattern observed when CoA is tested against peptide as variable substrate and at fixed concentrations of acetyl-CoA also rules out a ping-pong mechanism
CoA-SASEA
CoA covalently linked to substrate peptide fragment Ser1-Ala2-Ser3-Glu4-Ala5
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desulfo-CoA
-
competitive versus acetyl-CoA
diethyldicarbonate
-
-
iodoacetamide
-
-
iodoacetic acid
N-bromosuccinimide
-
-
N-ethylmaleimide
-
-
NH4+
-
partially inhibitory
p-chloromercuribenzoate
-
-
Ser-Tyr
-
inhibits NH2-terminal acetylation of adrenocorticotropic hormone (1-13)NH2
siRNA
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
-
cysteine
-
-
dithiothreitol
-
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glutathione
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.005 - 0.467
acetyl-CoA
0.0022 - 0.008
acetylCoA
0.096 - 0.276
adrenocorticotropic hormone (1-10)
-
0.035 - 0.16
adrenocorticotropic hormone (1-13)NH2
-
0.024
adrenocorticotropic hormone (1-16)NH2
-
-
-
0.011
adrenocorticotropic hormone (1-18)
-
-
-
0.0042 - 0.2
adrenocorticotropic hormone (1-24)
-
0.05 - 0.286
adrenocorticotropic hormone (1-39)
-
0.037
adrenocorticotropic hormone (4-10)NH2
-
-
-
0.043
beta-endorphin(1-27)
-
-
0.065
beta-endorphin(1-31)
-
-
0.3 - 1.85
SASEAGVRWGRPVGRRRRP
-
additional information
additional information
-
steady-state kinetics, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.48 - 0.58
acetyl-CoA
0.025 - 4.4
SASEAGVRWGRPVGRRRRP
-
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)
-
-
0.113
adrenocorticotropic hormone (1-2)[ser-tyr]
-
-
-
0.106
adrenocorticotropic hormone (1-24)
-
-
-
0.253
alpha-N,O-Diacetyl-alphaMSH
-
-
0.00227 - 0.00277
CoA
0.067
desulfo-CoA
-
with acetyl-CoA, pH 7.4, temperature not specified in the publication
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38
acetonyl-CoA
Schizosaccharomyces pombe
O74985
pH 8.0, 25C
-
0.0014
CoA-SASEA
Schizosaccharomyces pombe
O74985
pH 8.0, 25C
-
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
-
-
7.4 - 8
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assay at
8.3
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2 - 9.8
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
activity assay
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
strong expression
Manually annotated by BRENDA team
-
higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
-
primary thyroid cells
Manually annotated by BRENDA team
-
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
-
higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
-
higher hNAA16 than hNAA15 expression
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
-
-
Manually annotated by BRENDA team
additional information
endogenous hNaa40p localizes to both the cytoplasm and the nuclei
-
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
170000
-
-
180000
-
gel filtration
190000
-
gel filtration
240000
-
gel filtration
241000
-
holoenzyme, calculated on the basis of the subunits
250000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterotrimer
trimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetylation
isoform hARD1235 shows autoacetylation
acylation
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Naa50p bound to a native substrate peptide fragment and CoA, hanging drop vapor diffusion method, mixing of 12 mg/ml protein in 25 mM HEPES, pH 7.5, 100 mM NaCl, and 10mM dithiothreitol, with peptide MLGPEGGRWG solution, and crystallization solution, containing 16% PEG 8000, 20% glycerol, and 40 mM potassium phosphate, pH 5.0, in a 1:3:3 molar ratio, 20C, 1-3 days, X-ray diffraction structure determination and analysis at 2.75 A resolution
-
structure of the 100 kDa holo-NatA complex in the absence and presence of a bisubstrate peptide-CoA conjugate inhibitor, and structure of the uncomplexed Naa10p catalytic subunit. The NatA-Naa15p auxiliary subunit contains 13 TPR motifs and adopts a ring-like topology that wraps around the NatA-Naa10p subunit, an interaction that alters the Naa10p active site for substrate-specific acetylation
the purified protein is crystallized in complex with CoA and with CoA/peptide by sitting-drop vapour-diffusion method at 10C. 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 SsArd1CoA/peptide ternary complex at 1.84 A
-
hanging drop vapour-diffusion method, Ta0058 protein is crystallized at 24C 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 24C 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
-
about 25% maximal activity at pH 6.0 and 8.0
487649
7.6
-
pronounced decline in reaction rate at pH values above
487650
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
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irreversible denaturation after 1 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme is completely inactivated by lyophilization
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greatly stablized by inclusion of EDTA and 0.01% deoxycholate in the isolation buffer
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loses activity by digestion with bovine pancreatic RNase A, Staphylococcus aureus nuclease or proteinase K
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, enzyme is completely inactivated by freezing
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-20C, stored as Percoll fraction, activity declines by about 50% in 5 days
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-70C, 10-20% glycerol, activity is stable for at least 5 months
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-70C, stored as Percoll fraction, activity declines by about 50% in 5 days
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0-4C, 10% of the activity is lost after storage for a week
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0C, sensitive to freezing, more than 90% loss of activity per freeze-thaw cycle
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4C, 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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4C, stored as Percoll fraction, activity declines by about 50% in 5 days
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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 GST-tagged full-length Naa50p from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography and cleavage of the GST-tag by TEV protease, followed by ion exchange chromatography and gel filtration
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recombinant His-tagged and maltose binding protein-tagged subunits hNaa15p, hNaa10p, and hNaa50p from Escherichia coli by nickel affinity chromatography
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recombinant His-tagged subunits hNaa10p and hNaa50p from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
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recombinant hNaa40p-V5 from HEK293 cells by immunoprecipitation
ribosomes of HEK-293 cells are isolated
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3 N-terminal acetyltransferases, Ard1p/Nat1p3, Nat3p and Mak3p
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a plasmid expressing hNaa16p, NARG1L-FLAG, is used
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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
cloning of V5-tagged hNaa15p, hNaa25p, and hNaa35p from HEK-293 cell genomic DNA, expression of His-tagged and maltose binding protein-tagged subunits hNaa15p, hNaa10p, and hNaa50p in Escherichia coli, efficient expression of MBP-hNaa15p requires coexpression of pDC952, a plasmid carrying the Escherichia coli argU gene
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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 in Saccharomyces cerevisiae lacking endogenous Naa50
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expression of GST-tagged full-length Naa50p with a TEV protease-cleavable site in Escherichia coli strain BL21(DE3)
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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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recombinant expression of hNaa40p in Escherichia coli strain BL21 StarTM (DE3), and of hNaa40p-V5 in HEK293 cells. Heterologous ectopically expression of hNaa40p in a Saccharomyces cerevisiae naa40-DELTA strain restores Nt-acetylation of yeast histone H4, but not H2A in vivo
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
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F27A
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site-directed mutagenesis, inactive mutant
F35A
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site-directed mutagenesis, inactive mutant
H112F
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site-directed mutagenesis, inactive mutant
I142A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
P28A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
V29A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y139A
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site-directed mutagenesis, inactive mutant
Y31A
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site-directed mutagenesis, inactive mutant
Y73A
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site-directed mutagenesis, inactive mutant
Y73F
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site-directed mutagenesis, inactive mutant
E24A
mutation in NatA, decrease in kcat, increase in Km value
E24D
mutation in NatA, decrease in kcat, increase in Km value
E24Q
mutation in NatA, decrease in kcat, increase in Km value
E61A
mutation in NatA, decrease in kcat, increase in Km value
E62A
mutation in NatA, decrease in kcat, increase in Km value
H111A
mutation in NatA, decrease in kcat, Km value similar to wild-type
H20A
mutation in NatA, decrease in kcat, increase in Km value
H72A
mutation in NatA, decrease in kcat, increase in Km value
K29A
mutation in NatA, increase in kcat, decrease in Km value
K29A/Y33A
mutation in NatA, decrease in kcat, Km value similar to wild-type
K59A
mutation in NatA, decrease in kcat, increase in Km value
K59A/E61A
mutation in NatA, decrease in kcat, increase in Km value
K59A/E62A
mutation in NatA, decrease in kcat, increase in Km value
L22A
mutation in NatA, decrease in kcat, increase in Km value
P23A
mutation in NatA, decrease in kcat, increase in Km value
R113A
mutation in NatA, strong decrease in kcat, Km value similar to wild-type
R80A
mutation in NatA, decrease in kcat, increase in Km value
Y139A
mutation in NatA, dramaitc loss of activity
Y26A
mutation in NatA, decrease in kcat, increase in Km value
Y33A
mutation in NatA, decrease in kcat, increase in Km value
E35A
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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
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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
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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
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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
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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
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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
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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
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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)
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E35V
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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
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N132A
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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)
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R100A
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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
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T105A
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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)
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additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
development of a proteolysis-based assay, named PROMPT, i.e. PROtease assay for Membrane Protein Topology, to determine the topology of protein C-termini
medicine
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Naa50p is a therapeutic anti-cancer target, the structure of the ternary Naa50p complex also provides a molecular scaffold for the design of NAT-specific small molecule inhibitors with possible therapeutic applications
Show AA Sequence (147 entries)
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