EzCatDB: D00508

DB codeD00508
RLCP classification3.1107.220190.41
CATH domainDomain 12.40.128.40Catalytic domain
Domain 23.-.-.-
E.C.2.3.1.118
CSA1e2t


Enzyme Name
Swiss-protKEGG

Q00267
Protein nameN-hydroxyarylamine O-acetyltransferaseN-hydroxyarylamine O-acetyltransferase
arylhydroxamate N,O-acetyltransferase
arylamine N-acetyltransferase
N-hydroxy-2-aminofluorene-O-acetyltransferase
SynonymsEC 2.3.1.118
Arylhydroxamate N,O-acetyltransferase
Arylamine N-acetyltransferase
NAT101


Swiss-prot:Accession NumberQ00267
Entry nameNHOA_SALTY
ActivityAcetyl-CoA + an N-hydroxyarylamine = CoA + an N-acetoxyarylamine.
SubunitMonomer and homodimer.
Subcellular location
Cofactor


SubstratesProducts
KEGG-idC00024C02720C00010C02709
CompoundAcetyl-CoAN-HydroxyarylamineCoAN-Acetoxyarylamine
Typeamine group,carbohydrate,nucleotide,peptide/protein,sulfide groupamine group,aromatic ring (only carbon atom)amine group,carbohydrate,nucleotide,peptide/protein,sulfhydryl groupamine group,aromatic ring (only carbon atom),carbohydrate
1e2tA01UnboundUnboundUnboundUnbound
1e2tB01UnboundUnboundUnboundUnbound
1e2tC01UnboundUnboundUnboundUnbound
1e2tD01UnboundUnboundUnboundUnbound
1e2tE01UnboundUnboundUnboundUnbound
1e2tF01UnboundUnboundUnboundUnbound
1e2tG01UnboundUnboundUnboundUnbound
1e2tH01UnboundUnboundUnboundUnbound
1e2tA02UnboundUnboundUnboundUnbound
1e2tB02UnboundUnboundUnboundUnbound
1e2tC02UnboundUnboundUnboundUnbound
1e2tD02UnboundUnboundUnboundUnbound
1e2tE02UnboundUnboundUnboundUnbound
1e2tF02UnboundUnboundUnboundUnbound
1e2tG02UnboundUnboundUnboundUnbound
1e2tH02UnboundUnboundUnboundUnbound

Active-site residues
resource
PDB;1e2t & Swiss-prot;Q00267 & literature [17]
pdbCatalytic residuesMain-chain involved in catalysis
1e2tA01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tB01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tC01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tD01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tE01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tF01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tG01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tH01CYS 69;HIS 107;ASP 122
CYS 69;PHE 70
1e2tA02

1e2tB02

1e2tC02

1e2tD02

1e2tE02

1e2tF02

1e2tG02

1e2tH02


References for Catalytic Mechanism
ReferencesSectionsNo. 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]
PubMed ID6644748
JournalJ Med Chem
Year1983
Volume26
Pages1780-4
AuthorsBanks RB, Smith TJ, Hanna PE
TitleN-arylhydroxamic acid N,O-acyltransferase. Positional requirements for the substrate hydroxyl group
[2]
PubMed ID4045921
JournalJ Med Chem
Year1985
Volume28
Pages1453-60
AuthorsElfarra AA, Hanna PE
TitleSubstituent effects on the bioactivation of 2-(N-hydroxyacetamido)fluorenes by N-arylhydroxamic acid N,O-acyltransferase
[3]
PubMed ID3965709
JournalJ Med Chem
Year1985
Volume28
Pages18-24
AuthorsMarhevka VC, Ebner NA, Sehon RD, Hanna PE
TitleMechanism-based inactivation of N-arylhydroxamic acid N,O-acyltransferase by 7-substituted-N-hydroxy-2-acetamidofluorenes
[4]
PubMed ID3745141
JournalJ Biochem (Tokyo)
Year1986
Volume99
Pages1689-97
AuthorsSaito K, Shinohara A, Kamataki T, Kato R
TitleN-hydroxyarylamine O-acetyltransferase in hamster liver
[5]
PubMed ID2725469
JournalMol Pharmacol
Year1989
Volume35
Pages599-609
AuthorsMattano SS, Land S, King CM, Weber WW
TitlePurification and biochemical characterization of hepatic arylamine N-acetyltransferase from rapid and slow acetylator mice
[6]
PubMed ID2253236
JournalCancer Res
Year1990
Volume50
Pages7942-9
AuthorsTrinidad A, Hein DW, Rustan TD, Ferguson RJ, Miller LS, Bucher KD, Kirlin WG, Ogolla F, Andrews AF
TitlePurification of hepatic polymorphic arylamine N-acetyltransferase from homozygous rapid acetylator inbred hamster
[7]
PubMed ID1464118
JournalChem Pharm Bull (Tokyo)
Year1992
Volume40
Pages2857-9
AuthorsSone T, Yamaguchi T, Isobe M, Takabatake E, Adachi T, Hirano K, Wang CY
TitlePurification and characterization of hamster hepatic microsomal N,O-acetyltransferase
[8]
PubMed ID1569093
JournalJ Biol Chem
Year1992
Volume267
Pages8429-36
AuthorsWatanabe M, Sofuni T, Nohmi T
TitleInvolvement 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
[9]
PubMed ID8179482
JournalArch Toxicol
Year1994
Volume68
Pages129-33
AuthorsHein DW, Rustan TD, Ferguson RJ, Doll MA, Gray K
TitleMetabolic activation of aromatic and heterocyclic N-hydroxyarylamines by wild-type and mutant recombinant human NAT1 and NAT2 acetyltransferases
[10]
PubMed ID7889864
JournalEnviron Health Perspect
Year1994
Volume102 Suppl 6
Pages83-9
AuthorsWatanabe M, Igarashi T, Kaminuma T, Sofuni T, Nohmi T
TitleN-hydroxyarylamine O-acetyltransferase of Salmonella typhimurium
[11]
PubMed ID9535705
JournalProtein Expr Purif
Year1998
Volume12
Pages371-80
AuthorsSinclair JC, Delgoda R, Noble ME, Jarmin S, Goh NK, Sim E
TitlePurification, characterization, and crystallization of an N-hydroxyarylamine O-acetyltransferase from Salmonella typhimurium
[12]
PubMed ID10806332
JournalBiochim Biophys Acta
Year2000
Volume1475
Pages10-6
AuthorsYamamura E, Sayama M, Kakikawa M, Mori M, Taketo A, Kodaira K
TitlePurification and biochemical properties of an N-hydroxyarylamine O-acetyltransferase from Escherichia coli
[13]
CommentsX-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS), AND SUBUNIT STRUCTURE.
Medline ID20336895
PubMed ID10876241
JournalNat Struct Biol
Year2000
Volume7
Pages560-4
AuthorsSinclair JC, Sandy J, Delgoda R, Sim E, Noble ME
TitleStructure of arylamine N-acetyltransferase reveals a catalytic triad
Related PDB1e2t
Related Swiss-protQ00267
[14]
PubMed ID11368758
JournalBiochem J
Year2001
Volume356
Pages327-34
AuthorsRodrigues-Lima F, Delomenie C, Goodfellow GH, Grant DM, Dupret JM
TitleHomology 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.
[15]
PubMed ID11829470
JournalBiochem Biophys Res Commun
Year2002
Volume291
Pages116-23
AuthorsRodrigues-Lima F, Dupret JM
Title3D 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.
[16]
PubMed ID11799105
JournalJ Biol Chem
Year2002
Volume277
Pages12175-81
AuthorsMushtaq A, Payton M, Sim E
TitleThe COOH terminus of arylamine N-acetyltransferase from Salmonella typhimurium controls enzymic activity.
[17]
PubMed ID12054803
JournalJ Mol Biol
Year2002
Volume318
Pages1071-83
AuthorsSandy J, Mushtaq A, Kawamura A, Sinclair J, Sim E, Noble M
TitleThe structure of arylamine N-acetyltransferase from Mycobacterium smegmatis--an enzyme which inactivates the anti-tubercular drug, isoniazid.
[18]
PubMed ID11812235
JournalProtein Expr Purif
Year2002
Volume24
Pages138-51
AuthorsPompeo F, Mushtaq A, Sim E
TitleExpression and purification of the rifamycin amide synthase, RifF, an enzyme homologous to the prokaryotic arylamine N-acetyltransferases.
[19]
PubMed ID12595067
JournalBiochim Biophys Acta
Year2003
Volume1620
Pages8-14
AuthorsDelgoda R, Lian LY, Sandy J, Sim E
TitleNMR investigation of the catalytic mechanism of arylamine N-acetyltransferase from Salmonella typhimurium.
[20]
PubMed ID12628650
JournalBioorg Med Chem
Year2003
Volume11
Pages1227-34
AuthorsBrooke EW, Davies SG, Mulvaney AW, Pompeo F, Sim E, Vickers RJ
TitleAn approach to identifying novel substrates of bacterial arylamine N-acetyltransferases.
[21]
PubMed ID12852958
JournalBioorg Med Chem Lett
Year2003
Volume13
Pages2527-30
AuthorsBrooke EW, Davies SG, Mulvaney AW, Okada M, Pompeo F, Sim E, Vickers RJ, Westwood IM
TitleSynthesis and in vitro evaluation of novel small molecule inhibitors of bacterial arylamine N-acetyltransferases (NATs).

comments
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.
(5) His107 acts as a general base to activate the acceptor group of the second substrate, N-hydroxyarylamine.
(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.

createdupdated
2004-03-232009-02-26


Copyright: Nozomi Nagano, JST & CBRC-AIST
Funded by PRESTO/Japan Science and Technology Corporation (JST) (December 2001 - November 2004)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
Funded by Grant-in-Aid for Scientific Research (B)/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2010)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2011 - March 2012)

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