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Literature summary extracted from

  • Liszczak, G.; Arnesen, T.; Marmorsteins, R.
    Structure of a ternary Naa50p (NAT5/SAN) N-terminal acetyltransferase complex reveals the molecular basis for substrate-specific acetylation (2011), J. Biol. Chem., 286, 37002-37010.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
2.3.1.258 medicine 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 Homo sapiens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.258 expressed in Escherichia coli BL21(DE3) cells Homo sapiens
2.3.1.258 expression of GST-tagged full-length Naa50p with a TEV protease-cleavable site in Escherichia coli strain BL21(DE3) Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.3.1.258 enzyme bound to a native substrate peptide fragment (MLGPEGGRWG) and CoA, hanging drop vapor diffusion method, using 16% (w/v) PEG 8000, 20% (v/v) glycerol, and 40 mM potassium phosphate (monobasic, pH 5.0) Homo sapiens
2.3.1.258 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, 20°C, 1-3 days, X-ray diffraction structure determination and analysis at 2.75 A resolution Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.258 F27A site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 F27A the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens
2.3.1.258 F35A site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 F35A the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens
2.3.1.258 H112A site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 H112A the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens
2.3.1.258 H112F site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 H112F the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens
2.3.1.258 I142A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
2.3.1.258 I142A the mutant shows 42.2% of wild type catalytic efficiency Homo sapiens
2.3.1.258 P28A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
2.3.1.258 P28A the mutant shows less 13.4% of wild type catalytic efficiency Homo sapiens
2.3.1.258 V29A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
2.3.1.258 V29A the mutant shows 12.3% of wild type catalytic efficiency Homo sapiens
2.3.1.258 Y139A site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 Y139A the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens
2.3.1.258 Y31A site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 Y31A the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens
2.3.1.258 Y73A site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 Y73A the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens
2.3.1.258 Y73F site-directed mutagenesis, inactive mutant Homo sapiens
2.3.1.258 Y73F the mutant shows less than 10% of wild type catalytic efficiency Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.258 acetyl-CoA + MLGPEGGRWG Homo sapiens
-
CoA + Ac-MLGPEGGRWG
-
?
2.3.1.258 acetyl-CoA + peptide Homo sapiens
-
CoA + Nalpha-acetylpeptide
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.258 Homo sapiens
-
-
-
2.3.1.258 Homo sapiens Q9GZZ1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.258 HiTrap SP column chromatography, and Superdex 75 gel filtration Homo sapiens
2.3.1.258 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 Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.258 acetyl-CoA + MLGPEGGRWG
-
Homo sapiens CoA + Ac-MLGPEGGRWG
-
?
2.3.1.258 acetyl-CoA + MLGPEGGRWGRPVGRRRRP
-
Homo sapiens acetyl-CoA + Nalpha-acetyl-MLGPEGGRWGRPVGRRRRP
-
?
2.3.1.258 acetyl-CoA + peptide
-
Homo sapiens CoA + Nalpha-acetylpeptide
-
?
2.3.1.258 acetyl-CoA + peptide peptide substrate binding structure, overview Homo sapiens CoA + Nalpha-acetylpeptide
-
?

Synonyms

EC Number Synonyms Comment Organism
2.3.1.258 Naa50p
-
Homo sapiens
2.3.1.258 Naa50p subunit of NatA Homo sapiens
2.3.1.258 Naa50p (NAT5/SAN) N-terminal acetyltransferase complex
-
Homo sapiens
2.3.1.258 NAT
-
Homo sapiens
2.3.1.258 NAT5/SAN
-
Homo sapiens
2.3.1.258 NatA
-
Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.258 8
-
assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
2.3.1.258 acetyl-CoA
-
Homo sapiens