EzCatDB: D00508
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DB codeD00508
RLCP classification3.1107.220190.41 : Transfer
CATH domainDomain 12.40.128.40 : LipocalinCatalytic domain
Domain 23.-.-.-
E.C.2.3.1.118
CSA1e2t


Enzyme Name
UniProtKBKEGG

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
RefSeqNP_460541.1 (Protein)
NC_003197.1 (DNA/RNA sequence)
PfamPF00797 (Acetyltransf_2)
[Graphical view]


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

Compound table: links to PDB-related databases & PoSSuM

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
ChEBI15351
28902
15346

PubChem6302
444493
7518
87642
6816
153367
            
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 UniProtKBQ00267
[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


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