Literature summary for 2.3.1.57 extracted from

Filippova, E.V.; Weigand, S.; Osipiuk, J.; Kiryukhina, O.; Joachimiak, A.; Anderson, W.F.
Substrate-induced allosteric change in the quaternary structure of the spermidine N-acetyltransferase SpeG (2015), J. Mol. Biol., 427, 3538-3553.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme, dodecameric structure in a ligand-free form in three different conformational states, open, intermediate and closed, sitting drop vapor diffusion method, for open state crystals: mixing of 400 nl of 10 mg/ml protein in 100 mM sodium chloride, 10 mM HEPES, pH 7.5, with 400 nl of reservoir solution containing 8% isopropanol and 0.1 M Tris-HCl, pH 8.5, for closed state crystals: mixing of 0.001 ml of 8.5 mg/ml protein in 500 mM sodium chloride, 5 mM 2-mercaptoethanol, 10 mM Tris-HCl, pH 8.3, with 0.001 ml of reservoir solution that contains 0.05 M ammonium sulfate, 0.1 M tri-sodium citrate and 15% polyethylene glycol 8000, and for intermediate state crystals: 0.001 ml of 8.5 mg/ml protein in 500 mM sodium chloride, 5 mM 2-mercaptoethanol, 10 mM Tris-HCl, pH 8.3, and 0.001 ml of reservoir solution containing 0.01 M calcium chloride, 20% methanol, and 0.1 M Tris-HCl, pH 8.5, 19°C, X-ray diffraction structure determination and analysis at 2.38-2.88 A reolution, molecular replacement. All structures are crystallized in C2 space group symmetry and contain six monomers in the asymmetric unit cell. Two hexamers related by crystallographic 2fold symmetry form the SpeG dodecamer. The open and intermediate states have a unique open dodecameric ring	Vibrio cholerae

Crystallization (Comment)

Organism

purified enzyme, dodecameric structure in a ligand-free form in three different conformational states, open, intermediate and closed, sitting drop vapor diffusion method, for open state crystals: mixing of 400 nl of 10 mg/ml protein in 100 mM sodium chloride, 10 mM HEPES, pH 7.5, with 400 nl of reservoir solution containing 8% isopropanol and 0.1 M Tris-HCl, pH 8.5, for closed state crystals: mixing of 0.001 ml of 8.5 mg/ml protein in 500 mM sodium chloride, 5 mM 2-mercaptoethanol, 10 mM Tris-HCl, pH 8.3, with 0.001 ml of reservoir solution that contains 0.05 M ammonium sulfate, 0.1 M tri-sodium citrate and 15% polyethylene glycol 8000, and for intermediate state crystals: 0.001 ml of 8.5 mg/ml protein in 500 mM sodium chloride, 5 mM 2-mercaptoethanol, 10 mM Tris-HCl, pH 8.3, and 0.001 ml of reservoir solution containing 0.01 M calcium chloride, 20% methanol, and 0.1 M Tris-HCl, pH 8.5, 19°C, X-ray diffraction structure determination and analysis at 2.38-2.88 A reolution, molecular replacement. All structures are crystallized in C2 space group symmetry and contain six monomers in the asymmetric unit cell. Two hexamers related by crystallographic 2fold symmetry form the SpeG dodecamer. The open and intermediate states have a unique open dodecameric ring

Vibrio cholerae

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	allosteric substrate binding, bireactant random steady-state kinetic mechanism	Vibrio cholerae

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
220000	230000	ligand-free enzyme, gel filtration and small-angle X-ray scattering analysis	Vibrio cholerae
250000	-	polyamine-bound enzyme, gel filtration and small-angle X-ray scattering analysis	Vibrio cholerae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
3 acetyl-CoA + 2 spermine	Vibrio cholerae	-	3 CoA + N1-acetylspermine + N1,N12-diacetylspermine	-	?
3 acetyl-CoA + 2 spermine	Vibrio cholerae N16961	-	3 CoA + N1-acetylspermine + N1,N12-diacetylspermine	-	?
3 acetyl-CoA + spermidine	Vibrio cholerae	-	3 CoA + N1,N4,N8-triacetylspermidine	-	?
3 acetyl-CoA + spermidine	Vibrio cholerae N16961	-	3 CoA + N1,N4,N8-triacetylspermidine	-	?

Organism

Organism	UniProt	Comment	Textmining
Vibrio cholerae	Q9KL03	serotype O1, gene speG	-
Vibrio cholerae N16961	Q9KL03	serotype O1, gene speG	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
3 acetyl-CoA + 2 spermine	-	Vibrio cholerae	3 CoA + N1-acetylspermine + N1,N12-diacetylspermine	-	?
3 acetyl-CoA + 2 spermine	-	Vibrio cholerae N16961	3 CoA + N1-acetylspermine + N1,N12-diacetylspermine	-	?
3 acetyl-CoA + spermidine	-	Vibrio cholerae	3 CoA + N1,N4,N8-triacetylspermidine	-	?
3 acetyl-CoA + spermidine	-	Vibrio cholerae N16961	3 CoA + N1,N4,N8-triacetylspermidine	-	?
additional information	substrate-induced allosteric change of quaternary structure of spermidine N-acetyltransferase SpeG, overview	Vibrio cholerae	?	-	?
additional information	substrate-induced allosteric change of quaternary structure of spermidine N-acetyltransferase SpeG, overview	Vibrio cholerae N16961	?	-	?

Subunits

Subunits	Comment	Organism
dodecamer	substrate-induced allosteric change of quaternary structure of spermidine N-acetyltransferase SpeG, overview. The enzyme possesses unique open dodecameric state exists in solution	Vibrio cholerae

Synonyms

Synonyms	Comment	Organism
SpeG	-	Vibrio cholerae
spermidine N-acetyltransferase	-	Vibrio cholerae

Cofactor

Cofactor	Comment	Organism	Structure
acetyl-CoA	-	Vibrio cholerae

General Information

General Information	Comment	Organism
evolution	the enzyme is a member of the Gcn5-related N-acetyltransferase superfamily. The open and intermediate states of ligand-free enzyme have a unique open dodecameric ring. The SpeG dodecamer is asymmetric except for the one 2fold axis and is unlike any known dodecameric structure. The SpeG dodecamer is conserved in different bacterial species, structure analysis and comparisons, overview	Vibrio cholerae
physiological function	enzyme SpeG has an allosteric polyamine-binding site and acetylating polyamines regulate their intracellular concentrations	Vibrio cholerae