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Information on EC 2.3.1.259 - N-terminal methionine Nalpha-acetyltransferase NatF and Organism(s) Homo sapiens and UniProt Accession Q9H7X0
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2.3.1.259 N-terminal methionine Nalpha-acetyltransferase NatFIUBMB Comments
N-terminal-acetylases (NATs) catalyse the covalent attachment of an acetyl moiety from acetyl-CoA to the free alpha-amino group at the N-terminus of a protein. This irreversible modification neutralizes the positive charge at the N-terminus, makes the N-terminal residue larger and more hydrophobic, and prevents its removal by hydrolysis. NatF is found only in higher eukaryotes, and is absent from yeast. Unlike other Nat systems the enzyme is located in the Golgi apparatus. It faces the cytosolic side of intracellular membranes, and specifically acetylates transmembrane proteins whose N termini face the cytosol. NatF targets N-terminal L-methionine residues attached to Lys, Ser, Val, Leu, Gln, Ile, Tyr and Thr residues.
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The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
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an N-terminal-L-methionyl-[transmembrane protein]
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an N-terminal-Nalpha-acetyl-L-methionyl-[transmembrane protein]
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Synonyms
naa60, naa60p, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Naa60
Naa60
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-
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SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:N-terminal-Met-Lys/Ser/Val/Leu/Gln/Ile/Tyr/Thr-[transmembrane protein] Met-Nalpha-acetyltransferase
N-terminal-acetylases (NATs) catalyse the covalent attachment of an acetyl moiety from acetyl-CoA to the free alpha-amino group at the N-terminus of a protein. This irreversible modification neutralizes the positive charge at the N-terminus, makes the N-terminal residue larger and more hydrophobic, and prevents its removal by hydrolysis. NatF is found only in higher eukaryotes, and is absent from yeast. Unlike other Nat systems the enzyme is located in the Golgi apparatus. It faces the cytosolic side of intracellular membranes, and specifically acetylates transmembrane proteins whose N termini face the cytosol. NatF targets N-terminal L-methionine residues attached to Lys, Ser, Val, Leu, Gln, Ile, Tyr and Thr residues.
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
acetyl-CoA + an N-terminal-L-methionyl-[transmembrane protein]
an N-terminal-Nalpha-acetyl-L-methionyl-[transmembrane protein] + CoA
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-
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?
acetyl-CoA + MAPLDLDRWGRPVGRRRRPVRVYP
Nalpha-acetyl-MAPLDLDRWGRPVGRRRRPVRVYP + CoA
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-
-
?
acetyl-CoA + MDELFPLRWGRPVGRRRRPVRVYP
Nalpha-acetyl-MDELFPLRWGRPVGRRRRPVRVYP + CoA
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-
-
?
acetyl-CoA + MKKSYSGRWGRPVGRRRRPVRVYP
Nalpha-acetyl-MKKSYSGRWGRPVGRRRRPVRVYP + CoA
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-
-
?
acetyl-CoA + MLGPEGGRWGRPVGRRRRPVRVYP
Nalpha-acetyl-MLGPEGGRWGRPVGRRRRPVRVYP + CoA
second highest N-terminal acetylation activity
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-
?
acetyl-CoA + MLGTEGGRWGRPVGRRRRPVRVYP
Nalpha-acetyl-MLGTEGGRWGRPVGRRRRPVRVYP + CoA
highest N-terminal acetylation activity
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-
?
acetyl-CoA + an N-terminal-L-methionyl-[transmembrane protein]
an N-terminal-Nalpha-acetyl-L-methionyl-[transmembrane protein] + CoA
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-
-
-
?
additional information
?
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almost no activity with peptides SESSSKSRWGRPVGRRRRPVRVYP and SYSMDHFRWGRPVGRRRRPVRVYP
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?
additional information
?
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almost no activity with peptides SESSSKSRWGRPVGRRRRPVRVYP and SYSMDHFRWGRPVGRRRRPVRVYP
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-
?
additional information
?
-
the enzyme also harbors lysine Nepsilon-acetyltransferase activity to catalyze the acetylation of lysine epsilon-amine
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?
additional information
?
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the enzyme also harbors lysine Nepsilon-acetyltransferase activity to catalyze the acetylation of lysine epsilon-amine
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?
additional information
?
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the enzyme has a preference for acetylating N-termini of the transmembrane proteins
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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
acetyl-CoA + an N-terminal-L-methionyl-[transmembrane protein]
an N-terminal-Nalpha-acetyl-L-methionyl-[transmembrane protein] + CoA
-
-
-
?
acetyl-CoA + an N-terminal-L-methionyl-[transmembrane protein]
an N-terminal-Nalpha-acetyl-L-methionyl-[transmembrane protein] + CoA
-
-
-
-
?
additional information
?
-
the enzyme has a preference for acetylating N-termini of the transmembrane proteins
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-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.001
CoA-Ac-MAPLDLD-NH2
Homo sapiens
pH and temperature not specified in the publication
0.00244
CoA-Ac-MKAVQAD-NH2
Homo sapiens
pH and temperature not specified in the publication
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Naa60 attaches to membranes of the Golgi apparatus. Molecular determinants for the interaction of Naa60 with the membrane by means of computational modeling, in vitro liposome assays including intrinsic tryptophan fluorescence, circular dichroism, and isothermal titration calorimetry (ITC), and mutational analysis combined with localization determination in cellulo. Model of Naa60 membrane association and N-terminal acetylation at the membrane, overview. Peripheral binding to Golgi-like liposomes. Naa60 has a preference for phosphatidylinositol 4-phosphate which may recruit it to the Golgi membrane mainly through Arg212 and Arg215 of Pred-alpha2, and through Lys233 and Arg240 of the C-terminal tail
additional information
the C-terminal tail of Naa60 (amino acids 182-242), which constitutes an extra segment, not present in the other catalytic Naas, is defined as the part providing organellar localization. Human eGFP-tagged Naa60 displays an organellar localization recombinantly expressed in Saccharomyces cerevisiae, and this localization pattern is lost and replaced by a cytosolic signal upon truncation of the last 61 amino acids. Naa60 uses a membrane targeting mechanism that is also functional in fungi, although Naa60 is not found in this kingdom. After incubation of the Naa60(189-242) peptide with Golgi-like liposomes composed of 52% phosphatidylcholine, 20% phosphatidylethanolamine, 15% cholesterol, 8% phosphatidylinositol 4-phosphate, and 5% phosphatidylserine, a clear blue shift from 355 to 348 nm is observed, hence indicating an interaction between Naa60(189-242) and these liposomes, residue Trp227 dips into the membrane
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isoform Naa60 faces the cytosolic side of intracellular membranes. Localization depends on a C-terminal membrane-integrating region
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
Nalpha-acetyltransferase 60 (Naa60) belongs to a family of enzymes known as N-terminal acetyltransferases (NATs)
physiological function
additional information
catalytic mechanism of Naa60, structure-function analysis, molecular dynamics simulations, overview
physiological function
the enzyme is important for normal chromosome segregation
physiological function
enzyme Naa60 has importance in maintaining the structural integrity of the Golgi apparatus in human cells
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). NAA60_HUMAN
242
0
27451
Swiss-Prot
other Location (Reliability: 2)
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 3% (w/v) tacsimate, 0.1 M bis-Tris (pH 6.5), and 16% polyethylene glycol (PEG) 3350
in complex with acetyl-coenzyme A or coenzyme A, hanging drop vapor diffusion method, using 10 mM Tris pH 8.0, 75 mM NaCl, 0.5% (v/v) glycerol, 3% (v/v) Tacsimate pH 4.0, and 7.5% (w/v) polyethylene glycol 3350
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D81A
the mutant shows 136.9% catalytic efficiency compared to the wild type enzyme
D81A APPLICATION
the mutant shows about 2fold increased activity compared to the wild type enzyme
D83A
the mutant shows about 2.8fold increased activity compared to the wild type enzyme
E80A
the mutant shows about 3.6fold increased activity compared to the wild type enzyme
H138A
I167A
the mutant shows an approximate 2.5 to 4fold reduction in kcat and a small increase in KM value, so the catalytic efficiency of this mutant is approximately 3fold of the wild type enzyme
I190A/L191A/Y193A/I194A
site-directed mutagenesis, enzyme deletions suggest membrane interaction by two regions matching the predicted helices (Pred), hydrophobic amino acids in Pred-alpha are mutated. The I190A/L191A/Y193A/I194A mutation, which involves two of the buried amino acids from Pred-alpha1, causes a mixed organellar and cytosolic localization in contrast to the wild-type. This mutation either causes only partially impaired membrane integration of Pred-alpha1 or additional segments are able to maintain some membrane association without Pred-alpha1. Truncation and deletion constructs of eGFP-tagged Naa60 demonstrate regions in the C-terminal tail important for membrane association in cellulo
I36A
the mutant shows a 6fold reduction in catalytic efficiency compared to the wild type enzyme
I84A
the mutant shows 200.4% catalytic efficiency compared to the wild type enzyme
K79A
the mutant shows about 20% reduced activity compared to the wild type enzyme
K79Q
the mutant shows about 1.8fold increased activity compared to the wild type enzyme
K79R
the mutant shows about 1.5fold increased activity compared to the wild type enzyme
L140A
L197A/L201A/L204A/I209A
site-directed mutagenesis, the mutation results in a mixed localization of the enzyme, with a partial and almost completely extractable organellar fraction, but with a more severe degree of mislocalization compared with mutant I190A/L191A/Y193A/I194A
N143A
the mutant shows about 60% reduced activity compared to the wild type enzyme
P35A
the mutant shows a 6fold reduction in catalytic efficiency compared to the wild type enzyme
Y164A
the mutant shows 141.7% catalytic efficiency compared to the wild type enzyme
Y164F
the mutant shows 154.9% catalytic efficiency compared to the wild type enzyme
Y165F
the mutant has an approximately 4fold reduction in catalytic efficiency compared to the wild type enzyme
Y38A
Y97F
additional information
generation of truncated enzyme fragment Naa60(189-242), which peripherally binds Golgi-like liposomes possibly with a horizontal orientation of helices relative to the membrane bilayer
L140A
the mutant has a 4-5fold increase in kcat but at the same time a 5-10fold increase in KM value , so the catalytic efficiency of this mutant is approximately 0.5fold of the wild type enzyme
L140A
the mutant shows about 20% reduced activity compared to the wild type enzyme
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene NAA60, recombinant expression of eGFP-tagged enzyme in Saccharomyces cerevisiae in the Golgi apparatus
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
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
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Aksnes, H.; Van Damme, P.; Goris, M.; Starheim, K.K.; Marie, M.; St?ve, S.I.; Hoel, C.; Kalvik, T.V.; Hole, K.; Glomnes, N.; Furnes, C.; Ljostveit, S.; Ziegler, M.; Niere, M.; Gevaert, K.; Arnesen, T.
An organellar Nalpha-acetyltransferase, Naa60, acetylates cytosolic N termini of transmembrane proteins and maintains Golgi integrity
Cell Rep.
10
1362-1374
2015
Homo sapiens (Q9H7X0)
Aksnes, H.; Marie, M.; Arnesen, T.
Holding it together: Naa60 at the Golgi
Oncotarget
6
15726-15727
2015
Homo sapiens
Van Damme, P.; Hole, K.; Pimenta-Marques, A.; Helsens, K.; Vandekerckhove, J.; Martinho, R.G.; Gevaert, K.; Arnesen, T.
NatF contributes to an evolutionary shift in protein N-terminal acetylation and is important for normal chromosome segregation
PLoS Genet.
7
e1002169
2011
Homo sapiens (Q9H7X0), Homo sapiens
Chen, J.Y.; Liu, L.; Cao, C.L.; Li, M.J.; Tan, K.; Yang, X.; Yun, C.H.
Structure and function of human Naa60 (NatF), a Golgi-localized bi-functional acetyltransferase
Sci. Rep.
6
31425
2016
Homo sapiens (Q9H7X0), Homo sapiens
Osberg, C.; Aksnes, H.; Ninzima, S.; Marie, M.; Arnesen, T.
Microscopy-based Saccharomyces cerevisiae complementation model reveals functional conservation and redundancy of N-terminal acetyltransferases
Sci. Rep.
6
31627
2016
Homo sapiens
Stove, S.; Magin, R.; Foyn, H.; Haug, B.; Marmorstein, R.; Arnesen, T.
Crystal structure of the Golgi-associated human Nalpha-acetyltransferase 60 reveals the molecular determinants for substrate-specific acetylation
Structure
24
1044-1056
2016
Homo sapiens (Q9H7X0), Homo sapiens
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