Literature summary for 2.5.1.16 extracted from

Wu, H.; Min, J.; Ikeguchi, Y.; Zeng, H.; Dong, A.; Loppnau, P.; Pegg, A.E.; Plotnikov, A.N.
Structure and mechanism of spermidine synthases (2007), Biochemistry, 46, 8331-8339.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the His-tagged enzyme in Escherichia coli strain BL21 (DE3)	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme free or in complex with spermidine, putrescine, 5'-methyl, 5'-deoxymethylthioadenosine, and S-adenosylmethionine, hanging drop vapor diffusion method, 20°C, 10 mg/ml protein solution is mixed with reservoir solution, containing 20-25% PEG 3350, 0.1 M (NH4)2SO4, 0.1 M HEPES-NaOH, pH 7.5, and ligands in a ratio of 1: 5 enzyme-ligand, X-ray diffraction structure determination and analysis at 1.9-2.1 A resolution	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant enzyme free or in complex with spermidine, putrescine, 5'-methyl, 5'-deoxymethylthioadenosine, and S-adenosylmethionine, hanging drop vapor diffusion method, 20°C, 10 mg/ml protein solution is mixed with reservoir solution, containing 20-25% PEG 3350, 0.1 M (NH4)2SO4, 0.1 M HEPES-NaOH, pH 7.5, and ligands in a ratio of 1: 5 enzyme-ligand, X-ray diffraction structure determination and analysis at 1.9-2.1 A resolution

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
D101I	site-directed mutagenesis, almost inactive mutant	Thermotoga maritima
D170A	site-directed mutagenesis, the mutsnt shows reduced activity compared to the wild-type enzyme	Thermotoga maritima
D173A	site-directed mutagenesis, the mutsnt shows reduced activity compared to the wild-type enzyme	Thermotoga maritima
Y76F	site-directed mutagenesis, the mutsnt shows reduced activity compared to the wild-type enzyme	Thermotoga maritima

Inhibitors

Inhibitors	Comment	Organism	Structure
S-Adenosyl-1,8-diamino-3-thiooctane	-	Thermotoga maritima

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	knetics of wild-type and mutant enzymes	Thermotoga maritima
0.0075	-	S-adenosylmethioninamine	pH 7.5, 37°C, recombinant enzyme	Thermotoga maritima
0.009	-	S-adenosylmethioninamine	pH 7.5, 37°C, recombinant enzyme	Homo sapiens
0.019	-	putrescine	pH 7.5, 37°C, recombinant enzyme	Thermotoga maritima
0.02	-	putrescine	pH 7.5, 37°C, recombinant enzyme	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
S-adenosylmethioninamine + putrescine	Homo sapiens	-	5'-S-methyl-5'-thioadenosine + spermidine	-	?
S-adenosylmethioninamine + putrescine	Thermotoga maritima	-	5'-S-methyl-5'-thioadenosine + spermidine	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P19623	-	-
Thermotoga maritima	Q9WZC2	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography	Homo sapiens

Reaction

Reaction	Comment	Organism	Reaction ID
S-adenosyl 3-(methylsulfanyl)propylamine + putrescine = S-methyl-5'-thioadenosine + spermidine	general mechanistic hypothesis for the aminopropyl transfer reaction	Homo sapiens	a
S-adenosyl 3-(methylsulfanyl)propylamine + putrescine = S-methyl-5'-thioadenosine + spermidine	general mechanistic hypothesis for the aminopropyl transfer reaction	Thermotoga maritima	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
5'-methyl,5'-deoxymethylthioadenosine + putrescine	low activity	Homo sapiens	?	-	?
5'-methyl,5'-deoxymethylthioadenosine + putrescine	low activity	Thermotoga maritima	?	-	?
additional information	amine substrate specificity, overview	Homo sapiens	?	-	?
additional information	amine substrate specificity, overview	Thermotoga maritima	?	-	?
S-adenosylmethioninamine + putrescine	-	Homo sapiens	5'-S-methyl-5'-thioadenosine + spermidine	-	?
S-adenosylmethioninamine + putrescine	-	Thermotoga maritima	5'-S-methyl-5'-thioadenosine + spermidine	-	?
S-adenosylmethioninamine + putrescine	the enzyme is highly specific for putrescine	Homo sapiens	5'-S-methyl-5'-thioadenosine + spermidine	-	?
S-adenosylmethioninamine + putrescine	the enzyme shows a relatively low substrate specificity for putrescine	Thermotoga maritima	5'-S-methyl-5'-thioadenosine + spermidine	-	?
S-adenosylmethioninamine + putrescine	low activity	Homo sapiens	spermidine + 5'-methylthioadenosine	-	?
S-adenosylmethioninamine + putrescine	low activity	Thermotoga maritima	spermidine + 5'-methylthioadenosine	-	?

Subunits

Subunits	Comment	Organism
More	structure analysis, comparison to the enzyme from Thermotoga maritima	Homo sapiens
More	structure analysis, comparison to the human enzyme	Thermotoga maritima

Synonyms

Synonyms	Comment	Organism
SPDSYN	-	Homo sapiens
SPDSYN	-	Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens
37	80	assay at	Thermotoga maritima

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
37	80	-	Thermotoga maritima
37	50	-	Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
80	-	2 min, purified recombinant enzyme, stable	Thermotoga maritima
80	-	2 min, recombinant enzyme, inactivation	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
additional information	-	additional information	-	Thermotoga maritima
0.012	-	putrescine	pH 7.5, 80°C, recombinant mutant D101I	Thermotoga maritima
0.012	-	S-adenosylmethioninamine	pH 7.5, 80°C, recombinant mutant D101I	Thermotoga maritima
0.014	-	putrescine	pH 7.5, 80°C, recombinant mutant D170A	Thermotoga maritima
0.014	-	S-adenosylmethioninamine	pH 7.5, 80°C, recombinant mutant D170A	Thermotoga maritima
0.77	-	putrescine	pH 7.5, 37°C, recombinant wild-type enzyme	Thermotoga maritima
0.77	-	S-adenosylmethioninamine	pH 7.5, 37°C, recombinant wild-type enzyme	Thermotoga maritima
1.5	-	putrescine	pH 7.5, 80°C, recombinant mutant D173A	Thermotoga maritima
1.5	-	S-adenosylmethioninamine	pH 7.5, 80°C, recombinant mutant D173A	Thermotoga maritima
1.9	-	putrescine	pH 7.5, 37°C, recombinant enzyme	Homo sapiens
1.9	-	S-adenosylmethioninamine	pH 7.5, 37°C, recombinant enzyme	Homo sapiens
8.1	-	putrescine	pH 7.5, 80°C, recombinant mutant Y76F	Thermotoga maritima
8.1	-	S-adenosylmethioninamine	pH 7.5, 80°C, recombinant mutant Y76F	Thermotoga maritima
22.7	-	putrescine	pH 7.5, 80°C, recombinant wild-type enzyme	Thermotoga maritima
22.7	-	S-adenosylmethioninamine	pH 7.5, 80°C, recombinant wild-type enzyme	Thermotoga maritima

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Homo sapiens