Literature summary for 2.3.1.1 extracted from

Zhao, G.; Jin, Z.; Allewell, N.M.; Tuchman, M.; Shi, D.
Structures of the N-acetyltransferase domain of Xylella fastidiosa N-acetyl-L-glutamate synthase/kinase with and without a His tag bound to N-acetyl-L-glutamate (2015), Acta Crystallogr. Sect. F, 71, 86-95.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Xylella fastidiosa
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	Xylella fastidiosa

Crystallization (Commentary)

Crystallization (Comment)	Organism
enzyme bound to N-acetyl-L-glutamate, sitting drop vapor diffusion method, using 0.2 M Li2SO4, 0.1 M Tris pH 6.5, 25% (w/v) PEG 3350	Xylella fastidiosa
purified recombinant N-acetyltransferase domain of N-acetyl-L-glutamate synthase/kinase, with and without a His-tag, complexed with N-acetyl-L-glutamate, sitting-drop vapor-diffusion method, 0.002 ml of protein in 50 mM Tris-HCl, pH 7.4, 50 mM NaCl, 10% glycerol, 5 mM 2-mercaptoethanol, 1 mM EDTA, 10 mM CoA, and 10 mM N-acetyl-L-glutamate, is mixed with 0.2 M Li2SO4, 0.1 M Tris, pH 6.5, 25% PEG 3350 for the His-tagged enzyme, and with 0.2 M Li2SO4, 0.1 M Tris, pH 8.5, 25% PEG 3350 for the detagged enzyme, X-ray diffraction structure determination and analysis at 1.7 A and 1.4 A resolution, respectively	Xylella fastidiosa

Crystallization (Comment)

Organism

enzyme bound to N-acetyl-L-glutamate, sitting drop vapor diffusion method, using 0.2 M Li2SO4, 0.1 M Tris pH 6.5, 25% (w/v) PEG 3350

Xylella fastidiosa

purified recombinant N-acetyltransferase domain of N-acetyl-L-glutamate synthase/kinase, with and without a His-tag, complexed with N-acetyl-L-glutamate, sitting-drop vapor-diffusion method, 0.002 ml of protein in 50 mM Tris-HCl, pH 7.4, 50 mM NaCl, 10% glycerol, 5 mM 2-mercaptoethanol, 1 mM EDTA, 10 mM CoA, and 10 mM N-acetyl-L-glutamate, is mixed with 0.2 M Li2SO4, 0.1 M Tris, pH 6.5, 25% PEG 3350 for the His-tagged enzyme, and with 0.2 M Li2SO4, 0.1 M Tris, pH 8.5, 25% PEG 3350 for the detagged enzyme, X-ray diffraction structure determination and analysis at 1.7 A and 1.4 A resolution, respectively

Xylella fastidiosa

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
37400	-	catalytic N-acetyltransferase domain, gel filtration	Xylella fastidiosa
37400	-	recombinant dimeric N-acetyltransferase domain, gel filtration	Xylella fastidiosa
38800	-	catalytic N-acetyltransferase domain, calculated from amino acid sequence	Xylella fastidiosa

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
acetyl-CoA + L-glutamate	Xylella fastidiosa	-	CoA + N-acetyl-L-glutamate	-	?
acetyl-CoA + L-glutamate	Xylella fastidiosa ATCC 35881D	-	CoA + N-acetyl-L-glutamate	-	?
additional information	Xylella fastidiosa	the bifunctional N-acetyl-L-glutamate synthase/kinase contains an N-terminal catalytic N-acetyltransferase (NAT) domain, substrate N-acetyl-l-glutamate binding structure analysis, overview	?	-	?
additional information	Xylella fastidiosa ATCC 35881D	the bifunctional N-acetyl-L-glutamate synthase/kinase contains an N-terminal catalytic N-acetyltransferase (NAT) domain, substrate N-acetyl-l-glutamate binding structure analysis, overview	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Xylella fastidiosa	-	-	-
Xylella fastidiosa	Q87EL2	-	-
Xylella fastidiosa ATCC 35881D	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
HiTrap column chromatography	Xylella fastidiosa
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and cation exchange chromatography, the His-tag is cleaved by thrombin	Xylella fastidiosa

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
acetyl-CoA + L-glutamate	-	Xylella fastidiosa	CoA + N-acetyl-L-glutamate	-	?
acetyl-CoA + L-glutamate	-	Xylella fastidiosa ATCC 35881D	CoA + N-acetyl-L-glutamate	-	?
additional information	the bifunctional N-acetyl-L-glutamate synthase/kinase contains an N-terminal catalytic N-acetyltransferase (NAT) domain, substrate N-acetyl-l-glutamate binding structure analysis, overview	Xylella fastidiosa	?	-	?
additional information	the bifunctional N-acetyl-L-glutamate synthase/kinase contains an N-terminal catalytic N-acetyltransferase (NAT) domain, substrate N-acetyl-l-glutamate binding structure analysis, overview	Xylella fastidiosa ATCC 35881D	?	-	?

Subunits

Subunits	Comment	Organism
dimer	isolated N-acetyltransferase domain of N-acetyl-L-glutamate synthase/kinase	Xylella fastidiosa
homodimer	x-ray crystallography	Xylella fastidiosa
More	eight subunits of N-acetyltransferase domain form four molecular dimers in the crystal structure, overview	Xylella fastidiosa

Synonyms

Synonyms	Comment	Organism
N-acetyl-L-glutamate synthase/kinase	-	Xylella fastidiosa
N-acetyl-L-glutamate synthase/kinase	bifunctional enzyme	Xylella fastidiosa
NAGS	-	Xylella fastidiosa
NAGS/K	-	Xylella fastidiosa
NAT	catalytic N-acetyltransferase domain	Xylella fastidiosa

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Xylella fastidiosa

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.5	-	assay at	Xylella fastidiosa

Cofactor

Cofactor	Comment	Organism	Structure
acetyl-CoA	-	Xylella fastidiosa