DB code: D00508

RLCP classification 3.1107.220190.41 : Transfer
CATH domain 2.40.128.40 : Lipocalin Catalytic domain
3.-.-.- :
E.C. 2.3.1.118
CSA 1e2t
M-CSA 1e2t
MACiE

CATH domain Related DB codes (homologues)

Uniprot Enzyme Name
UniprotKB Protein name Synonyms RefSeq Pfam
Q00267 N-hydroxyarylamine O-acetyltransferase
EC 2.3.1.118
Arylhydroxamate N,O-acetyltransferase
Arylamine N-acetyltransferase
NAT101
NP_460541.1 (Protein)
NC_003197.1 (DNA/RNA sequence)
PF00797 (Acetyltransf_2)
[Graphical View]

KEGG enzyme name
N-hydroxyarylamine O-acetyltransferase
arylhydroxamate N,O-acetyltransferase
arylamine N-acetyltransferase
N-hydroxy-2-aminofluorene-O-acetyltransferase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
Q00267 NHOA_SALTY Acetyl-CoA + an N-hydroxyarylamine = CoA + an N-acetoxyarylamine. Monomer and homodimer.

KEGG Pathways
Map code Pathways E.C.

Compound table
Substrates Products Intermediates
KEGG-id C00024 C02720 C00010 C02709
E.C.
Compound Acetyl-CoA N-Hydroxyarylamine CoA N-Acetoxyarylamine
Type amine group,carbohydrate,nucleotide ,peptide/protein,sulfide group amine group,aromatic ring (only carbon atom) amine group,carbohydrate,nucleotide ,peptide/protein,sulfhydryl group amine group,aromatic ring (only carbon atom),carbohydrate
ChEBI 15351
 15351
 		28902
 28902
 		15346
 15346
PubChem 444493
 6302
 444493
 6302
 		7518
 7518
 		6816
 87642
 6816
 87642
 		153367
 153367
1e2tA01 Unbound Unbound Unbound Unbound
1e2tB01 Unbound Unbound Unbound Unbound
1e2tC01 Unbound Unbound Unbound Unbound
1e2tD01 Unbound Unbound Unbound Unbound
1e2tE01 Unbound Unbound Unbound Unbound
1e2tF01 Unbound Unbound Unbound Unbound
1e2tG01 Unbound Unbound Unbound Unbound
1e2tH01 Unbound Unbound Unbound Unbound
1e2tA02 Unbound Unbound Unbound Unbound
1e2tB02 Unbound Unbound Unbound Unbound
1e2tC02 Unbound Unbound Unbound Unbound
1e2tD02 Unbound Unbound Unbound Unbound
1e2tE02 Unbound Unbound Unbound Unbound
1e2tF02 Unbound Unbound Unbound Unbound
1e2tG02 Unbound Unbound Unbound Unbound
1e2tH02 Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
PDB;1e2t & Swiss-prot;Q00267 & literature [17]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1e2tA01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tB01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tC01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tD01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tE01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tF01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tG01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tH01 CYS 69;HIS 107;ASP 122 CYS 69;PHE 70
1e2tA02
1e2tB02
1e2tC02
1e2tD02
1e2tE02
1e2tF02
1e2tG02
1e2tH02

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[8]
Fig.8, p.8434-8435
[10]
Fig.4, p.86-87
[13]
p.561-563
[17]
p.1076-1078, p.1079

References
[1]
Resource
Comments
Medline ID
PubMed ID 6644748
Journal J Med Chem
Year 1983
Volume 26
Pages 1780-4
Authors Banks RB, Smith TJ, Hanna PE
Title N-arylhydroxamic acid N,O-acyltransferase. Positional requirements for the substrate hydroxyl group
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 4045921
Journal J Med Chem
Year 1985
Volume 28
Pages 1453-60
Authors Elfarra AA, Hanna PE
Title Substituent effects on the bioactivation of 2-(N-hydroxyacetamido)fluorenes by N-arylhydroxamic acid N,O-acyltransferase
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 3965709
Journal J Med Chem
Year 1985
Volume 28
Pages 18-24
Authors Marhevka VC, Ebner NA, Sehon RD, Hanna PE
Title Mechanism-based inactivation of N-arylhydroxamic acid N,O-acyltransferase by 7-substituted-N-hydroxy-2-acetamidofluorenes
Related PDB
Related UniProtKB
[4]
Resource
Comments
Medline ID
PubMed ID 3745141
Journal J Biochem (Tokyo)
Year 1986
Volume 99
Pages 1689-97
Authors Saito K, Shinohara A, Kamataki T, Kato R
Title N-hydroxyarylamine O-acetyltransferase in hamster liver
Related PDB
Related UniProtKB
[5]
Resource
Comments
Medline ID
PubMed ID 2725469
Journal Mol Pharmacol
Year 1989
Volume 35
Pages 599-609
Authors Mattano SS, Land S, King CM, Weber WW
Title Purification and biochemical characterization of hepatic arylamine N-acetyltransferase from rapid and slow acetylator mice
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 2253236
Journal Cancer Res
Year 1990
Volume 50
Pages 7942-9
Authors Trinidad A, Hein DW, Rustan TD, Ferguson RJ, Miller LS, Bucher KD, Kirlin WG, Ogolla F, Andrews AF
Title Purification of hepatic polymorphic arylamine N-acetyltransferase from homozygous rapid acetylator inbred hamster
Related PDB
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 1464118
Journal Chem Pharm Bull (Tokyo)
Year 1992
Volume 40
Pages 2857-9
Authors Sone T, Yamaguchi T, Isobe M, Takabatake E, Adachi T, Hirano K, Wang CY
Title Purification and characterization of hamster hepatic microsomal N,O-acetyltransferase
Related PDB
Related UniProtKB
[8]
Resource
Comments
Medline ID
PubMed ID 1569093
Journal J Biol Chem
Year 1992
Volume 267
Pages 8429-36
Authors Watanabe M, Sofuni T, Nohmi T
Title Involvement of Cys69 residue in the catalytic mechanism of N-hydroxyarylamine O-acetyltransferase of Salmonella typhimurium. Sequence similarity at the amino acid level suggests a common catalytic mechanism of acetyltransferase for S. typhimurium and higher organisms
Related PDB
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 8179482
Journal Arch Toxicol
Year 1994
Volume 68
Pages 129-33
Authors Hein DW, Rustan TD, Ferguson RJ, Doll MA, Gray K
Title Metabolic activation of aromatic and heterocyclic N-hydroxyarylamines by wild-type and mutant recombinant human NAT1 and NAT2 acetyltransferases
Related PDB
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 7889864
Journal Environ Health Perspect
Year 1994
Volume 102 Suppl 6
Pages 83-9
Authors Watanabe M, Igarashi T, Kaminuma T, Sofuni T, Nohmi T
Title N-hydroxyarylamine O-acetyltransferase of Salmonella typhimurium
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 9535705
Journal Protein Expr Purif
Year 1998
Volume 12
Pages 371-80
Authors Sinclair JC, Delgoda R, Noble ME, Jarmin S, Goh NK, Sim E
Title Purification, characterization, and crystallization of an N-hydroxyarylamine O-acetyltransferase from Salmonella typhimurium
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 10806332
Journal Biochim Biophys Acta
Year 2000
Volume 1475
Pages 10-6
Authors Yamamura E, Sayama M, Kakikawa M, Mori M, Taketo A, Kodaira K
Title Purification and biochemical properties of an N-hydroxyarylamine O-acetyltransferase from Escherichia coli
Related PDB
Related UniProtKB
[13]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS), AND SUBUNIT STRUCTURE.
Medline ID 20336895
PubMed ID 10876241
Journal Nat Struct Biol
Year 2000
Volume 7
Pages 560-4
Authors Sinclair JC, Sandy J, Delgoda R, Sim E, Noble ME
Title Structure of arylamine N-acetyltransferase reveals a catalytic triad
Related PDB 1e2t
Related UniProtKB Q00267
[14]
Resource
Comments
Medline ID
PubMed ID 11368758
Journal Biochem J
Year 2001
Volume 356
Pages 327-34
Authors Rodrigues-Lima F, Delomenie C, Goodfellow GH, Grant DM, Dupret JM
Title Homology modelling and structural analysis of human arylamine N-acetyltransferase NAT1: evidence for the conservation of a cysteine protease catalytic domain and an active-site loop.
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 11829470
Journal Biochem Biophys Res Commun
Year 2002
Volume 291
Pages 116-23
Authors Rodrigues-Lima F, Dupret JM
Title 3D model of human arylamine N-acetyltransferase 2: structural basis of the slow acetylator phenotype of the R64Q variant and analysis of the active-site loop.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 11799105
Journal J Biol Chem
Year 2002
Volume 277
Pages 12175-81
Authors Mushtaq A, Payton M, Sim E
Title The COOH terminus of arylamine N-acetyltransferase from Salmonella typhimurium controls enzymic activity.
Related PDB
Related UniProtKB
[17]
Resource
Comments
Medline ID
PubMed ID 12054803
Journal J Mol Biol
Year 2002
Volume 318
Pages 1071-83
Authors Sandy J, Mushtaq A, Kawamura A, Sinclair J, Sim E, Noble M
Title The structure of arylamine N-acetyltransferase from Mycobacterium smegmatis--an enzyme which inactivates the anti-tubercular drug, isoniazid.
Related PDB
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 11812235
Journal Protein Expr Purif
Year 2002
Volume 24
Pages 138-51
Authors Pompeo F, Mushtaq A, Sim E
Title Expression and purification of the rifamycin amide synthase, RifF, an enzyme homologous to the prokaryotic arylamine N-acetyltransferases.
Related PDB
Related UniProtKB
[19]
Resource
Comments
Medline ID
PubMed ID 12595067
Journal Biochim Biophys Acta
Year 2003
Volume 1620
Pages 8-14
Authors Delgoda R, Lian LY, Sandy J, Sim E
Title NMR investigation of the catalytic mechanism of arylamine N-acetyltransferase from Salmonella typhimurium.
Related PDB
Related UniProtKB
[20]
Resource
Comments
Medline ID
PubMed ID 12628650
Journal Bioorg Med Chem
Year 2003
Volume 11
Pages 1227-34
Authors Brooke EW, Davies SG, Mulvaney AW, Pompeo F, Sim E, Vickers RJ
Title An approach to identifying novel substrates of bacterial arylamine N-acetyltransferases.
Related PDB
Related UniProtKB
[21]
Resource
Comments
Medline ID
PubMed ID 12852958
Journal Bioorg Med Chem Lett
Year 2003
Volume 13
Pages 2527-30
Authors Brooke EW, Davies SG, Mulvaney AW, Okada M, Pompeo F, Sim E, Vickers RJ, Westwood IM
Title Synthesis and in vitro evaluation of novel small molecule inhibitors of bacterial arylamine N-acetyltransferases (NATs).
Related PDB
Related UniProtKB

Comments
(5) His107 acts as a general base to activate the acceptor group of the second substrate, N-hydroxyarylamine.
Although the CATH classification of this enzyme structure has not been determined yet, it is homologous to coagulation factor XIII A chain (E.C. 2.3.2.13, Swiss-prot; P00488, PDB;1f13), whose catalytic domain structure is classified into CATH 3.90.260.10. The catalytic residues are also conserved between these two enzymes.
According to the literature [13] & [17], the reaction proceeds as follows:
(1) Asp122 modulates the activity of His107.
(2) His107 acts as a general base to activate the sidechain Sgamma atom of Cys69.
(3) Cys69 makes a nucleophilic attack on the carbonyl carbon of Acetyl-CoA, leading to a tetrahedral transition-state. Here, the negatively charged transition-state is stabilized by an oxyanion hole, which is made up by mainchain amide groups of Cys69 and Phe70.
(4) His107 acts as a general acid to protonate the leaving group, sulfhydryl group of CoA. At the same time, the transition-state might collapse to form a covalent acyl-enzyme intermediate.
(6) The activated acceptor group makes a nucleophilic attack on the acyl-enzyme intermediate, leading to a tetrahedral transition-state. Here, the negatively charged transition-state is stabilized by the oxyanion hole.
(7) His107 acts as a general acid to protonate the leaving group, completing the reaction.

Created Updated
2004-03-23 2009-02-26