Information on EC 2.5.1.22 - spermine synthase and Organism(s) Homo sapiens

for references in articles please use BRENDA:EC2.5.1.22
Word Map on EC 2.5.1.22
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
This record set is specific for:
Homo sapiens


The taxonomic range for the selected organisms is: Homo sapiens

The enzyme appears in selected viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.5.1.22
-
RECOMMENDED NAME
GeneOntology No.
spermine synthase
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aminopropyl group transfer
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
spermine biosynthesis
-
-
superpathway of polyamine biosynthesis II
-
-
polyamine pathway
-
-
Cysteine and methionine metabolism
-
-
Arginine and proline metabolism
-
-
beta-Alanine metabolism
-
-
Glutathione metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl 3-(methylthio)propylamine:spermidine 3-aminopropyltransferase
The enzyme from mammalia is highly specific for spermidine [2,3]. cf. EC 2.5.1.16 (spermidine synthase) and EC 2.5.1.23 (sym-norspermidine synthase).
CAS REGISTRY NUMBER
COMMENTARY hide
74812-43-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
H2N(CH2)3NH(CH2)4NH2 + S-adenosyl 3-(methylthio)propylamine
H2N(CH2)3NH(CH2)4NH(CH2)3NH2 + S-methyl-5'-thioadenosine
show the reaction diagram
-
-
-
-
?
S-adenosyl 3-(methylthio)propylamine + spermidine
S-methyl-5'-thioadenosine + spermine
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methioninamine + spermidine
5'-methylthioadenosine + spermine
show the reaction diagram
S-adenosylmethioninamine + spermidine
5'-methylthioadenosine + spermine + H+
show the reaction diagram
-
Polyamine sythesis, addition of a second aminopropyl group to the N-10 position of spermidine. The active site with a bound spermidine molecule contains an Asp276 residue, which is in an ideal position to facilitate the deprotonation of the N-10 amino group of spermidine that attacks the C-atom of the aminopropyl group of decarboxylated S-adenosylmethionine
-
-
?
S-adenosylmethioninamine + spermidine
S-methyl-5'-thioadenosine + spermine
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl 3-(methylthio)propylamine + spermidine
S-methyl-5'-thioadenosine + spermine
show the reaction diagram
-
-
-
-
?
S-adenosylmethioninamine + spermidine
5'-methylthioadenosine + spermine + H+
show the reaction diagram
-
Polyamine sythesis, addition of a second aminopropyl group to the N-10 position of spermidine. The active site with a bound spermidine molecule contains an Asp276 residue, which is in an ideal position to facilitate the deprotonation of the N-10 amino group of spermidine that attacks the C-atom of the aminopropyl group of decarboxylated S-adenosylmethionine
-
-
?
S-adenosylmethioninamine + spermidine
S-methyl-5'-thioadenosine + spermine
show the reaction diagram
-
decarboxylated S-adenosylmethionine is an essential intermediate in the synthesis of polyamines
-
-
?
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5'-methylthioadenosine
-
SpmSyn is strongly inhibited by 5'-methylthioadenosine. This inhibition does not have great importance in limiting spermine synthesis in vivo because 5'-methylthioadenosine is normally rapidly degraded by 5'-methylthioadenosine phosphorylase. Inhibition of this enzyme allows 5'-methylthioadenosine to accumulate with deleterious effects on polyamine content.
additional information
-
the spermine synthase-5'-methylthioadenosine structure provides a plausible explanation for the potent inhibition of the reaction by this product and the stronger inhibition of spermine synthase compared with spermidine synthase.
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0005 - 0.0009
S-adenosyl 3-(methylthio)propylamine
0.001
S-adenosylmethioninamine
-
-
0.2 - 0.8
spermidine
additional information
additional information
-
ratio of kcat/Km value of wild-type is 40000 for substrate spermidine, and 71000000 for S-adenosyl-L-methioninamine, respectively
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0015 - 32
spermidine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0003
5'-methylthioadenosine
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.455
-
placenta
227
-
mutant enzyme S165D/L175E/T178H/C206R, at pH 7.5 and 37C
2425
-
wild type enzyme, at pH 7.5 and 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
additional information
-
tissue distribution
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
Homo sapiens;
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
-
2 * 45000, kidney, SDS-PAGE
78000
-
kidney, pore-gradient electrophoresis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
-
deletions of all or part of the N-terminal domain lead to the protein existing as a monomer as determined by gel filtration analysis and to virtually complete loss of activity
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
analysis of X-ray crystal structure of the wild-type human SMS in complex with spermidine and 5-methylthioadenosine, PDB ID 3C6K, and folding free energy calculations, dimer structure analysis of wild-type and mutant enzymes, overview
-
in complex with 5'-methylthioadenosine and spermidine and with 5'-methylthioadenosine and spermine. Enzyme is a dimer of two identical subunits. Each monomer has three domains, a C-terminal domain, which contains the active site and is similar in structure to spermidine synthase, a central domain made up of four beta-strands, and an N-terminal domain with remarkable structural similarity to S-adenosylmethionine decarboxylase
-
Purified SpmSyn is crystallized as ternary complex in the presence of spermidine and 5'-methylthioadenosine or spermidine and 5'-methylthioadenosine using the hanging drop vapor diffusion method. The SpmSyn-5'-methylthioadenosine-spermidine complex is crystallized in 12% polyethylene glycol and 0.1 M MES (pH 6.5). The SpmSyn-5'-methylthioadenosine-spermine complex is crystallized in 18% polyethylene glycol, 0.1 M NaCl, and 0.1 M BisTris (pH 6.5). Crystals are soaked in the corresponding mother liquor supplemented with 20% glycerol as cryoprotectant before freezing in liquid nitrogen.
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cell lysate is loaded onto a HiTrap chelating column charged with Ni2+. The enzyme is eluted with an imidazole gradient (50-250 mM) at pH 8.0. The protein is loaded onto a Superdex 200 column equilibrated with 20 mM Tris-HCl and 150 mM NaCl. Thrombin is added to combined fractions containing SpmSyn to remove the His-tag. The protein is further purified to homogeneity by ion-exchange chromatography.
-
TALON affinity resin column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA fragment encoding human enzyme is amplified by PCR and subcloned into the pET28a-LIC vector downstream of the polyhistidine coding region. Enzyme is expressed in the Escherichia coli BL21-Codon Plus(DE3)-RIL strain by the addition of 1 mM isopropyl1-thio-beta-D-galactopyranoside. The N-terminally truncated SpmSyn mutants are generated by PCR and subcloned into the pET28a-LIC vector.
-
enzyme overexpression in transgenic mice under control of a composite CMV-IE enhancer-chicken beta-actin promotor, 4 separate founder CAG/SpmS mice are analysed: enzyme expression in all tissues, mostly highly increased compared to the wild-type mice, overview
-
expressed in Escherichia coli XL-1 Blue cells
-
expression of human spermine synthase in CAGSMS line 8 mice from a composite CMV-IE enhancer/chicken beta-actin promoter. Transgenic expression of spermine synthase in the Gy mice reverse all of the increases in dcAdoMet content and AdoMetDC
-
expression of wild-type and mutant enzymes in HEK cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
DELTA1-129
-
0.02% activity compared to the wild-type enzyme
DELTA1-145
-
no activity
DELTA1-19
-
0.003% activity compared to the wild-type enzyme
DELTA1-43
-
0.0002% activity compared to the wild-type enzyme
DELTA1-82
-
0.00023% activity compared to the wild-type enzyme
DELTA347-366
-
truncation of the protein at position 346 removing the last 20 residues lead to a complete loss of activity
DELTA358-366A
-
smaller truncation of only 9 residues has a smaller effect but still reduced activity by 75%
F58L
-
the mutation is associated with the Snyder-Robinson syndrome
G67E
-
the mutation is associated with the Snyder-Robinson syndrome
M35R
-
the mutation is associated with the Snyder-Robinson syndrome
P112L
-
the mutation is associated with the Snyder-Robinson syndrome
S165D/L175E/T178H/C206R
-
the mutant shows increased activity compared to the wild type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine