Literature summary for 2.3.1.5 extracted from

Fullam, E.; Westwood, I.M.; Anderton, M.C.; Lowe, E.D.; Sim, E.; Noble, M.E.
Divergence of cofactor recognition across evolution: coenzyme A binding in a prokaryotic arylamine N-acetyltransferase (2008), J. Mol. Biol., 375, 178-191.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli as a His-tagged fusion protein	Mycobacterium marinum
into the pET28b+ vector for expression in Escherichia coli BL21DE3 pLysS cells	Mycobacterium marinum

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structure of MMNAT in complex with CoA is determined. The overall MMNAT structure consists of three domains. Domain I forms an alpha-helical bundle (amino acids 4-88), domain II forms a beta-barrel (amino acids 89-200) and domain III forms an alpha/beta lid (amino acids 201-275). Assembled together, the molecular surface of these domains presents a deep and wide active-site cleft, at the base of which is found the catalytic cysteine residue. Surprisingly, the principal CoA recognition site in MMNAT is located some 30 A from the site of CoA recognition in the deposited structure of human NAT2 bound to CoA	Mycobacterium marinum
the crystal structure of the native enzyme is presented at a resolution of 1.9 A, cocrystallized with CoA at 2.4 A	Mycobacterium marinum

Crystallization (Comment)

Organism

crystal structure of MMNAT in complex with CoA is determined. The overall MMNAT structure consists of three domains. Domain I forms an alpha-helical bundle (amino acids 4-88), domain II forms a beta-barrel (amino acids 89-200) and domain III forms an alpha/beta lid (amino acids 201-275). Assembled together, the molecular surface of these domains presents a deep and wide active-site cleft, at the base of which is found the catalytic cysteine residue. Surprisingly, the principal CoA recognition site in MMNAT is located some 30 A from the site of CoA recognition in the deposited structure of human NAT2 bound to CoA

Mycobacterium marinum

the crystal structure of the native enzyme is presented at a resolution of 1.9 A, cocrystallized with CoA at 2.4 A

Mycobacterium marinum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.2	-	5-Aminosalicylate	-	Mycobacterium marinum
0.2	-	acetyl-CoA	using 5-aminosalicylate as a substrate	Mycobacterium marinum

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium marinum	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
using a Ni-NTA resin	Mycobacterium marinum
using Ni-NTA chromatography	Mycobacterium marinum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	MMNAT is found to acetylate a broad range of substrates, including isoniazid, 4-anisidine and 4-aminoveratrole, compounds that have been identified as substrates of the NAT enzyme of Mycobacterium tuberculosis. The specific activity profile differs somewhat from that of the other characterised mycobacterial NAT, NAT from Mycobacterium smegmatis, particularly in the rate of acetylation of hydralazine and 2-aminofluorene, which are both acetylated 100times more rapidly by MMNAT	Mycobacterium marinum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
acetyl-CoA + 2-aminofluorene	-	Mycobacterium marinum	CoA + N-acetyl-2-aminofluorene	-	?
acetyl-CoA + 4-aminobenzoic acid	-	Mycobacterium marinum	CoA + N-acetyl-4-aminobenzoic acid	-	?
acetyl-CoA + 4-aminoveratrole	-	Mycobacterium marinum	CoA + N-acetyl-4-aminoveratrole	-	?
acetyl-CoA + 4-aminoveratrole	-	Mycobacterium marinum	?	-	?
acetyl-CoA + 4-anisidine	-	Mycobacterium marinum	CoA + N-acetyl-4-anisidine	-	?
acetyl-CoA + 4-bromoaniline	-	Mycobacterium marinum	CoA + N-(4-bromophenyl)acetamide	-	?
acetyl-CoA + 4-bromoaniline	-	Mycobacterium marinum	CoA + N-acetyl-4-bromoaniline	-	?
acetyl-CoA + 4-butoxyaniline	-	Mycobacterium marinum	CoA + N-acetyl-4-butoxyaniline	-	?
acetyl-CoA + 4-butoxyaniline	-	Mycobacterium marinum	?	-	?
acetyl-CoA + 4-chloroaniline	-	Mycobacterium marinum	CoA + N-acetyl-4-chloroaniline	-	?
acetyl-CoA + 4-chloroaniline	-	Mycobacterium marinum	CoA + N-(4-chlorophenyl)acetamide	-	?
acetyl-CoA + 4-chlorobenzoic hydrazide	-	Mycobacterium marinum	?	-	?
acetyl-CoA + 4-chlorobenzoic hydrazide	-	Mycobacterium marinum	CoA + N-acetyl-4-chlorobenzoic hydrazide	-	?
acetyl-CoA + 4-ethoxyaniline	-	Mycobacterium marinum	CoA + N-acetyl-4-ethoxyaniline	-	?
acetyl-CoA + 4-ethoxyaniline	-	Mycobacterium marinum	?	-	?
acetyl-CoA + 4-hexyloxyaniline	-	Mycobacterium marinum	CoA + N-acetyl-4-hexyloxyaniline	-	?
acetyl-CoA + 4-hexyloxyaniline	-	Mycobacterium marinum	?	-	?
acetyl-CoA + 4-iodoaniline	-	Mycobacterium marinum	CoA + N-acetyl-4-iodoaniline	-	?
acetyl-CoA + 4-iodoaniline	-	Mycobacterium marinum	CoA + N-(4-iodophenyl)acetamide	-	?
acetyl-CoA + 4-phenoxyaniline	-	Mycobacterium marinum	CoA + N-acetyl-4-phenoxyaniline	-	?
acetyl-CoA + 4-phenoxyaniline	-	Mycobacterium marinum	?	-	?
acetyl-CoA + 5-aminosalicylate	-	Mycobacterium marinum	CoA + N-acetyl-5-aminosalicylate	-	?
acetyl-CoA + hydralazine	-	Mycobacterium marinum	?	-	?
acetyl-CoA + hydralazine	-	Mycobacterium marinum	CoA + N-acetyl-hydralazine	-	?
acetyl-CoA + isoniazid	-	Mycobacterium marinum	CoA + N-acetyl-isoniazid	-	?
acetyl-CoA + isoniazid	-	Mycobacterium marinum	?	-	?
acetyl-CoA + p-aminobenzoic acid	-	Mycobacterium marinum	CoA + N-acetyl-4-aminobenzoic acid	-	?
acetyl-CoA + procainamide	-	Mycobacterium marinum	?	-	?
acetyl-CoA + procainamide	-	Mycobacterium marinum	CoA + N-acetyl-procainamide	-	?

Subunits

Subunits	Comment	Organism
dimer	crystal structure	Mycobacterium marinum

Synonyms

Synonyms	Comment	Organism
arylamine N-acetyltransferase	-	Mycobacterium marinum
MMNAT	-	Mycobacterium marinum
NAT	-	Mycobacterium marinum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
28	-	assay at	Mycobacterium marinum
28	-	activity assay	Mycobacterium marinum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Mycobacterium marinum
8	-	activity assay	Mycobacterium marinum

Cofactor

Cofactor	Comment	Organism	Structure
acetyl-CoA	-	Mycobacterium marinum