Literature summary for 2.3.1.5 extracted from

Martins, M.; Pluvinage, B.; de la Sierra-Gallay, I.L.; Barbault, F.; Dairou, J.; Dupret, J.M.; Rodrigues-Lima, F.
Functional and structural characterization of the arylamine N-acetyltransferase from the opportunistic pathogen Nocardia farcinica (2008), J. Mol. Biol., 383, 549-560.

Application

Application	Comment	Organism
medicine	there is increasing evidence that arylamine N-acetyltransferases could contribute to antibiotic resistance in pathogenic bacteria	Nocardia farcinica

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli as a His-tagged fusion protein	Nocardia farcinica
into the pET28a vector for expression in Escherichia coli BL21DE3 pLysS cells	Nocardia farcinica

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structure of NfNAT is solved at a resolution of 2.7 A. Despite low sequence identity, enzyme shares an almost identical fold with that of other prokaryotic NATs, e.g. Mycobacterium smegmatis NAT and Mycobacterium marinum NAT. The overall NfNAT structure consists of three domains of equivalent size. The first two domains, a helical bundle (amino acids 1-98) and a beta-barrel (amino acids 99-199), are disposed in such a way that three residues (Cys82, His119, and Asp136) form a catalytic triad. The third domain is linked to the second through an interdomain helix (alpha6: amino acids 200-210). As shown for other prokaryotic NAT structures, the interface formed between domains 2 and 3 forms a substantial active-site cleft	Nocardia farcinica
the structure of NfNAT is solved at a resolution of 2.7 A	Nocardia farcinica

Crystallization (Comment)

Organism

crystal structure of NfNAT is solved at a resolution of 2.7 A. Despite low sequence identity, enzyme shares an almost identical fold with that of other prokaryotic NATs, e.g. Mycobacterium smegmatis NAT and Mycobacterium marinum NAT. The overall NfNAT structure consists of three domains of equivalent size. The first two domains, a helical bundle (amino acids 1-98) and a beta-barrel (amino acids 99-199), are disposed in such a way that three residues (Cys82, His119, and Asp136) form a catalytic triad. The third domain is linked to the second through an interdomain helix (alpha6: amino acids 200-210). As shown for other prokaryotic NAT structures, the interface formed between domains 2 and 3 forms a substantial active-site cleft

Nocardia farcinica

the structure of NfNAT is solved at a resolution of 2.7 A

Nocardia farcinica

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
1.414	-	isoniazid	-	Nocardia farcinica
1.414	-	isoniazid	apparent KM	Nocardia farcinica

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
36000	-	SDS-PAGE	Nocardia farcinica
36000	-	determined by SDS-PAGE and Western Blot analysis	Nocardia farcinica

Organism

Organism	UniProt	Comment	Textmining
Nocardia farcinica	-	-	-
Nocardia farcinica	Q5YYQ3	-	-

Purification (Commentary)

Purification (Comment)	Organism
using HIS-Select resin	Nocardia farcinica

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.013	-	substrate: 2-aminofluorene	Nocardia farcinica
0.015	-	sulfamethazine	Nocardia farcinica
0.015	-	substrate: sulfamethazine	Nocardia farcinica
0.023	-	substrate: beta-naphthylamine	Nocardia farcinica
0.024	-	substrate: 4-aminobiphenyl	Nocardia farcinica
0.02463	-	substrate: 5-aminosalicylate	Nocardia farcinica
0.029	-	substrate: sulfamethoxazole	Nocardia farcinica
0.029	-	sulfamethoxazole	Nocardia farcinica
0.052	-	substrate: benzidine	Nocardia farcinica
0.079	-	substrate: 4-aminoveratrole	Nocardia farcinica
0.087	-	substrate: hydralazine	Nocardia farcinica
0.111	-	4-aminobenzoic acid	Nocardia farcinica
0.111	-	substrate: 4-aminobenzoic acid	Nocardia farcinica
0.372	-	4-aminosalicylate	Nocardia farcinica
0.372	-	substrate: 4-aminosalicylate	Nocardia farcinica
2.878	-	isoniazid	Nocardia farcinica
2.878	-	substrate: isoniazid	Nocardia farcinica
13.03	-	4-aminofluorene	Nocardia farcinica
23.73	-	beta-naphthylamine	Nocardia farcinica
24.63	-	5-aminosalicylate	Nocardia farcinica
24.91	-	2-aminobiphenyl	Nocardia farcinica
52.33	-	benzidine	Nocardia farcinica
79.39	-	4-aminoveratrole	Nocardia farcinica
87.76	-	hydralazine	Nocardia farcinica

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
acetyl-CoA + 2-aminobiphenyl	-	Nocardia farcinica	?	-	?
acetyl-CoA + 2-aminofluorene	-	Nocardia farcinica	CoA + N-acetyl-2-aminofluorene	-	?
acetyl-CoA + 4-aminobenzoic acid	-	Nocardia farcinica	CoA + N-acetyl-4-aminobenzoic acid	-	?
acetyl-CoA + 4-aminobenzoic acid	-	Nocardia farcinica	?	-	?
acetyl-CoA + 4-aminobiphenyl	-	Nocardia farcinica	CoA + N-acetyl-4-aminobiphenyl	-	?
acetyl-CoA + 4-aminofluorene	-	Nocardia farcinica	?	-	?
acetyl-CoA + 4-aminosalicylate	-	Nocardia farcinica	CoA + N-acetyl-4-aminosalicylate	-	?
acetyl-CoA + 4-aminosalicylate	-	Nocardia farcinica	?	-	?
acetyl-CoA + 4-aminoveratrole	-	Nocardia farcinica	CoA + N-acetyl-4-aminoveratrole	-	?
acetyl-CoA + 4-aminoveratrole	-	Nocardia farcinica	?	-	?
acetyl-CoA + 5-aminosalicylate	-	Nocardia farcinica	CoA + N-acetyl-5-aminosalicylate	-	?
acetyl-CoA + 5-aminosalicylate	-	Nocardia farcinica	?	-	?
acetyl-CoA + benzidine	-	Nocardia farcinica	?	-	?
acetyl-CoA + benzidine	-	Nocardia farcinica	CoA + N-acetyl-benzidine	-	?
acetyl-CoA + beta-naphthylamine	-	Nocardia farcinica	?	-	?
acetyl-CoA + beta-naphthylamine	-	Nocardia farcinica	CoA + N-acetyl-beta-naphthylamine	-	?
acetyl-CoA + hydralazine	-	Nocardia farcinica	?	-	?
acetyl-CoA + hydralazine	-	Nocardia farcinica	CoA + N-acetyl-hydralazine	-	?
acetyl-CoA + isoniazid	-	Nocardia farcinica	CoA + N-acetyl-isoniazid	-	?
acetyl-CoA + isoniazid	-	Nocardia farcinica	?	-	?
acetyl-CoA + sulfamethazine	-	Nocardia farcinica	?	-	?
acetyl-CoA + sulfamethazine	-	Nocardia farcinica	CoA + N-acetyl-sulfamethazine	-	?
acetyl-CoA + sulfamethoxazole	-	Nocardia farcinica	CoA + N-acetyl-sulfamethoxazole	-	?
acetyl-CoA + sulfamethoxazole	-	Nocardia farcinica	?	-	?

Synonyms

Synonyms	Comment	Organism
arylamine N-acetyltransferase	-	Nocardia farcinica
NAT	-	Nocardia farcinica
NfNAT	-	Nocardia farcinica

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Nocardia farcinica
37	-	activity assay	Nocardia farcinica

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Nocardia farcinica
7.5	-	activity assay	Nocardia farcinica

Cofactor

Cofactor	Comment	Organism	Structure
acetyl-CoA	-	Nocardia farcinica