Literature summary for 2.3.1.259 extracted from

Aksnes, H.; Goris, M.; Stroemland, O.; Drazic, A.; Waheed, Q.; Reuter, N.; Arnesen, T.
Molecular determinants of the N-terminal acetyltransferase Naa60 anchoring to the Golgi membrane (2017), J. Biol. Chem., 292, 6821-6837 .

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Cloned(Commentary)

Cloned (Comment)	Organism
gene NAA60, recombinant expression of eGFP-tagged enzyme in Saccharomyces cerevisiae in the Golgi apparatus	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
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	Homo sapiens
I209W/W227F	site-directed mutagenesis	Homo sapiens
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	Homo sapiens
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	Homo sapiens
Y193W/W227F	site-directed mutagenesis	Homo sapiens
Y214W/W227F	site-directed mutagenesis	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
Golgi membrane	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	Homo sapiens	139	-
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	Homo sapiens	-	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
acetyl-CoA + an N-terminal-L-methionyl-[transmembrane protein]	Homo sapiens	-	an N-terminal-Nalpha-acetyl-L-methionyl-[transmembrane protein] + CoA	-	?
additional information	Homo sapiens	the enzyme has a preference for acetylating N-termini of the transmembrane proteins	?	-	-

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	Q9H7X0	-	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
HeLa cell	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
acetyl-CoA + an N-terminal-L-methionyl-[transmembrane protein]	-	Homo sapiens	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	Homo sapiens	?	-	-

Synonyms

Synonyms	Comment	Organism
N-terminal acetyltransferase	-	Homo sapiens
Naa60	-	Homo sapiens
Nalpha-acetyltransferase 60	-	Homo sapiens
NatF	-	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
acetyl-CoA	-	Homo sapiens

General Information

General Information	Comment	Organism
evolution	Nalpha-acetyltransferase 60 (Naa60) belongs to a family of enzymes known as N-terminal acetyltransferases (NATs)	Homo sapiens
additional information	catalytic mechanism of Naa60, structure-function analysis, molecular dynamics simulations, overview	Homo sapiens
physiological function	enzyme Naa60 has importance in maintaining the structural integrity of the Golgi apparatus in human cells	Homo sapiens

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Release 2023.1 (January 2023)