Information on EC 3.6.4.3 - microtubule-severing ATPase

Word Map on EC 3.6.4.3
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:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.6.4.3
-
RECOMMENDED NAME
GeneOntology No.
microtubule-severing ATPase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + H2O = ADP + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase (tubulin-dimerizing)
Another member of the AAA-ATPase family, active in splitting microtubules into tubulin dimers in the centrosome.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Actinocoryne sp.
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
no activity in Saccharomyces cerevisiae
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Sea urchin
recombinant enzyme expressed in Baculovirus/Sf9 system
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme is a member of a family of AAA+ ATPases (ATPase associated with various cellular activities)
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
ATP + H2O
ADP + phosphate
show the reaction diagram
GTP + H2O
GDP + phosphate
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
ATP + H2O
ADP + phosphate
show the reaction diagram
GTP + H2O
GDP + phosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methanamine
-
-
2-(4-tert-butylphenyl)pyridine
-
-
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-amine
-
-
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-carbaldehyde
-
-
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-carbonitrile
-
-
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-ol
-
-
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl methyl sulfone
-
-
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl methyl sulfoxide
-
-
3'-fluoro-N,N-dimethyl-4'-(trifluoromethyl)biphenyl-4-sulfonamide
-
-
3'-fluoro-N-methyl-4'-(trifluoromethyl)biphenyl-4-sulfonamide
-
-
3-(4-tert-butylphenyl)pyridine
-
-
3-fluoro-4'-(methoxymethyl)-4-(trifluoromethyl)biphenyl
-
-
3-[[3-(methylsulfanyl)phenyl]amino]-4-[4-(trifluoromethyl)phenyl]-1H-pyrrole-2,5-dione
-
inhibits microtubule-stimulated ATP hydrolysis by KSP
4'-(ethylsulfonyl)-3-fluoro-4-(trifluoromethyl)biphenyl
-
-
4'-(trifluoromethyl)biphenyl-4-carbonitrile
-
-
4'-tert-butylbiphenyl-4-carbonitrile
-
-
4-(2-tert-butylphenyl)pyridine
-
-
4-(3-tert-butylphenyl)pyridine
-
-
4-(4-tert-butylphenyl)pyridine
-
-
5'-adenylyl-beta,gamma-imidodiphosphate
-
purley competitive inhibitor
ATPgammaS
Ca2+
inhibits microtuble severing activity
ispinesib
-
SB-715992
methyl 3'-fluoro-4'-(trifluoromethyl)biphenyl-4-carboxylate
-
-
N-[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methanesulfonamide
-
-
N-[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]sulfamide
-
-
N-[5-[3-fluoro-4-(trifluoromethyl)phenyl]pyridin-2-yl]sulfamide
-
-
N-[[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methyl]sulfamide
-
-
protein tau
-
affords protection of microtubules in katanin-overexpressing neurons
PVZB1084
-
-
PVZB1194
-
inhibition only in presence of microtubules
sb26
-
mutation in the tbb-2 beta-tubulin gene that partially inhibits MEI-1/MEI-2 activity
-
spg4atg1
morpholino antisense oligonucleotide for blocking translation of spg4 mRNA
-
spg4exon7
morpholino antisense oligonucleotide for blocking correct splicing of spg4 mRNA
-
Tau
-
affords some protection of microtubules in spastin-overexpressing neurons
-
tubulin
-
stimulation up to 0.001 mM, inhibition above
-
[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methanol
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-[[3-(methylsulfanyl)phenyl]amino]-4-[4-(trifluoromethyl)phenyl]-1H-pyrrole-2,5-dione
-
accelerates KSP-catalyzed ATP hydrolysis in the absence of microtubules circa 10fold
microtubule
-
microtubules
-
SPAS-1's ATPase activity increases with increasing concentrations of microtubules up to 0.00025 mM and then declines at higher microtubule concentrations. In the presence of 0.00025 mM microtubules, ATPase activity is 7times higher than in the absence of microtubules
-
Protein kinase C
-
adseverin upregulation is linked to the chondrocyte maturation program and is regulated by PKC
-
tubulin
-
at 0.001 mM, up to 6fold stimulation of ATPase activity
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.014 - 2.2
ATP
0.27
GST-SPG-4
-
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.005 - 13
ATP
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0012 - 0.006
3-[[3-(methylsulfanyl)phenyl]amino]-4-[4-(trifluoromethyl)phenyl]-1H-pyrrole-2,5-dione
0.11
ATPgammaS
-
pH 6.8, temperature not specified in the publication
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00247
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-amine
Homo sapiens
-
-
0.00147
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-carbaldehyde
Homo sapiens
-
-
0.0014
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-carbonitrile
Homo sapiens
-
-
0.00214
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-ol
Homo sapiens
-
-
0.00086
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl methyl sulfone
Homo sapiens
-
-
0.0037
3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl methyl sulfoxide
Homo sapiens
-
-
0.00461
3'-fluoro-N-methyl-4'-(trifluoromethyl)biphenyl-4-sulfonamide
Homo sapiens
-
-
0.0068
3-(4-tert-butylphenyl)pyridine
Homo sapiens
-
-
0.00714
3-fluoro-4'-(methoxymethyl)-4-(trifluoromethyl)biphenyl
Homo sapiens
-
-
0.00549
4'-(trifluoromethyl)biphenyl-4-carbonitrile
Homo sapiens
-
-
0.00147
4'-tert-butylbiphenyl-4-carbonitrile
Homo sapiens
-
-
0.00042
4-(3-tert-butylphenyl)pyridine
Homo sapiens
-
-
0.001
4-(4-tert-butylphenyl)pyridine
Homo sapiens
-
-
0.00278
N-[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methanesulfonamide
Homo sapiens
-
-
0.00002
N-[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]sulfamide
Homo sapiens
-
-
0.00004
N-[5-[3-fluoro-4-(trifluoromethyl)phenyl]pyridin-2-yl]sulfamide
Homo sapiens
-
-
0.00306
N-[[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methyl]sulfamide
Homo sapiens
-
-
0.0007
PVZB1084
Homo sapiens
-
-
0.00012
PVZB1194
Homo sapiens
-
-
0.00363
[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methanol
Homo sapiens
-
-
additional information
1-[3'-fluoro-4'-(trifluoromethyl)biphenyl-4-yl]methanamine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000045
-
His6-spastin
0.000046
-
GST-spastin
0.07
-
37°C, pH 8.8
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.9
-
ATPase activity assay
7.5
-
ATPase activity assay
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
microtubule severing assay
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
cytoskeleton at the perinuclear region
Manually annotated by BRENDA team
-
mostly in the soluble fraction after ultracentrifugation
-
Manually annotated by BRENDA team
-
synaptic terminal of neutrons
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28000
-
GST-AtKSS1, determined by SDS-PAGE
44000
-
full-length spastin without tag, determined by SDS-PAGE
46000
-
GST-AtKSS2, determined by SDS-PAGE
57000
-
SPAS-1L, consists of nine exons, determined by SDS-PAGE and Western blotting
60100
-
theoretical value, calculated from amino acid sequence
62200
-
spastin isoform: 1st ATG, predicted molecular weight of the isoform tagged with His6
66000
-
GST-AtKSS13, determined by SDS-PAGE
67200
-
isoform, first ATG, + exon 4
68000
-
Western blotting, isoform M1
69000
-
GST-spastin, determined by SDS-PAGE
82000
-
GST-AtKSS23, determined by SDS-PAGE
84000
x * 84000, deduced from genen sequence
85000
-
fusion protein: p60 plant homolog-RFP, SDS-PAGE, Western blot
88000
-
monomer, GST-spastin, determined by gel filtration analysis and Western blotting
95000
-
GST-spastin analyzed on a 12%-PAG
350000
-
hexamer, GST-free spastin, determined by gel filtration analysis and Western blotting
530000
-
hexamer, GST-spastin, determined by gel filtration analysis and Western blotting
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
hexamer
homohexamer
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
secondary structure analyzed by circular dichroism (CD) spectrum. SPAS-1 retains a high degree of alpha-helical structure
-
the X-ray structure of the nucleotide-free, monomeric AAA domain of Drosophila melanogaster spastin, residues 464-758, is solved at a resolution of 2.7 A
hanging drop method, the N-terminal residues (about 10000 Da) are clipped off during time of crystal formation (5-7 days)
-
the structure of kp60-NTD reveals a striking similarity to those of the microtubule interacting and trafficking domains (MIT). The arragnement of helices 2 and 3 is well conserved between kp60-NTD and the MIT domain from Vps4, a homologous protein that promotes disassembly of the endosomal sorting complexes required for transport III membrane skeleton complex. The positively charged surface formed by helices 2 and 3 binds tubulin
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
47
-
the melting temperature Tm is estimated from the denaturation curve
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
90% pure after Ni2+-NTA agarose, analyzed by SDS-PAGE
-
baculovirus-expressed, recombinant DM-Kat60
-
copurification of p81 and p60
-
GSH-Sepharose affinity chromatography, ion exchange chromatography, cleavage with precission protease, GSH-Sepharose affinity chromatography and gel filtration
-
GST tagged p60-katanin purified using glutathione-Sepharose
GSTrap column chromatography, GST tag cleaved with PreScission Protease, GST-Sepharose chromatography, gel filtration
-
Ni-NTA column, glutathione-agarose beads
-
p60, 60 kDa catalytic subunit
-
purification of recombinant protein using a glutathione-sepharose column, removing of the GST moiety with precision protease
-
recombinant glutathione S-transferase-tagged form kp60-NTD, affinity purification on glutathione-Sepharose and dialysis
-
recombinant His-tagged katanin p60 subunit from Escherichia coli strain BL21(DE3) pLysS by nickel affinity chromatography to over 95% purity
recombinant untagged short isoform of MEI-1 subunit and MBP-TEV-tagged MEI-2 subunit from Escherichia coli by amylose affinity chromatography, eluted with TEV protease
-
using a glutathione Sepharose 4B column
-
using a glutathione-agarose column, the glutathione-S-transferase moiety is cleaved with thrombin
-
using a glutathione-Sepharose column, cleavage of the GST moiety with PreScission Protease
-
using Ni-NTA agarose affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a glutathione S-transferase-tagged form of kp60-NTD expressed in Escherichia coli BL21 (DE3)
-
adaptor-p80 fragments expressed in Escherichia coli BL21(DE3), GST tagged p60-katanin expressed in Escherichia coli JM109
baculo system for pull-down assay, TNT in vitro translation kit for S35-Met-labeled recombinant protein, cloned into pEGFP or pcDNA3.1 for transfection of COS-7 cells for immuno precipitation
-
baculovirus-expressed
-
cDNA in TA cloning vector for sequencing, for expression in cultured neurons or fibroblasts in pEGFP and pCMV-Tag vector
-
cloned in pDsRed vector for transfection of chondrocytes
-
cloning of Cp60 katanin cDNA, expressed in Escherichia coli BL21(DE3)
-
coding sequence of spastin fused N-terminally to EGFP (pEGFP-N3 vector)
-
D-spastin-GFP for localization studies
-
expressed in 293T HEK cells
-
expressed in Escherichia coli BL21(DE3), FLAG tagged versions expressed in HEK293 cells
expressed in NIH-3T3 cells; spastin cDNA was cloned into pGEX6p-3 for production of recombinant protein in Escherichia coli or into pEYFP-C1
-
gene Katna1, cloning of the enzyme katanin p60 subunit from rat liver, recombinant expression of His-tagged protein in Escherichia coli strain BL21(DE3) pLysS
GST fusion of amino acid residues 228-616 containing the E442Q mutation (42500 Da for the 389 amino acid residues construct (amino acid residues 228-616)), expressed in Escherichia coli BL21-CodonPlus (DE3)-RIPL
-
His tagged variants and GST fusions expressed in Escherichia coli BL21 (RIL)
-
human cDNA PCR amplified
human katanin cloned and expressed in Escherichia coli
-
human spastin was cloned into the pGBK-T7 plasmid for yeast two-hybrid screening
-
into pQE30 vector, His-tag, and pGEX 4T3, GST-fusion, for expression in Escherichia coli
-
into the pGEX-6P-3 vector for expression in Escherichia coli BL21-CodonPlus DE3-RIPL cells
-
into the TOPO TA cloning vector, into the plasmid pPD49.26
-
isolation of cDNA clones for the p60 and p80 subunits, heterologous expression of p60 in Sf9 cell baculovirus system
-
Mei-1 and Mei-2 cloned and coexpressed in HeLa cells
-
pBSK vector for subcloning of the PCR products, pACN and pbinSRN for generating plasmids coding for fusion constructs of the target protein
-
recombinant expression of untagged short isoform of MEI-1 subunit, coexpression with MBP-TEV-tagged MEI-2 subunit, in Escherichia coli
-
SPAS-1 protein (451 amino acid reisdues) is produced as an N-terminally His6-tagged recombinant protein, expressed in Escherichia coli BL21(DE3)
-
spastin cDNA is cloned into the pGEMT easy vector, for generating a bait construct for yeast two-hybrid analysis, the excised spastin fragment is cloned into the pGBKT7 vector, for expression analysis in mammalian cells, the fragment is cloned into the pCS2-myc and pEGFP-C1 vectors
-
TA-cloned for sequencing, subcloned into pFLAG-CMV-6a for transfection of HEK293 cells
the human short isoform of spastin initiating from the alternative start codon is amplified by PCR and cloned into the pET41a vector for expression in Escherichia coli BL21DE3 cells
-
the motor domain of wild-type KSP monomer, aa 1-360, is expressed in Escherichia coli BL21DE3 cells
-
three domains of AtKSS are defined, AtKSS1, aa 1-34, AtKSS2, aa 35-210, AtKSS3, the ATPase domain, aa 211-523, in addition, combinations are defined, AtKKS12 and AtKKS23, AtKSS1,2,3,12,and 23 are cloned into pGEX-4T-1 and pOL-LT plasmids
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression and cellular distribution are increased in neoplastic glial phenotypes, especially in glioblastoma
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E278Q
-
Walker B mutant SPAS-1, abolished ATPase activity
K224R
-
Walker A mutant SPAS-1, abolished ATPase activity
K257A
-
exhibits 130% of wild-type ATPase activity
K437P/K441P
predicted not to form the C-terminal alpha-helix, microtuble network remains in mutant protein expressing HEK293 cells
D655N
-
disease mutation
E542A
-
disease mutation, Walker B motif
K488R
-
disease mutation, Walker A motif
S462C
-
disease mutation
1-279STOP
-
deletion mutant, no severing of microtubules, no binding of microtubules
1-328STOP
-
deletion mutant, no severing of microtubules, but binding of microtubules
406-415del
-
localizes as wild type, but lacks microtubule severing activity
Delta1-132
-
YFP-spastin mutant, amino acids 1-132 deleted
DELTA1-226
-
full severing activity
DELTA1-227
-
deletion mutant, severs microtubules, binds microtubules
Delta116-194
-
YFP-spastin mutant, amino acids 116-194 deleted
Delta195-227
-
YFP-spastin mutant, amino acids 195-227 deleted
DELTAMTBD
-
deletion mutant lacking aa 1-227 and 270-328, neither binds nor severs microtubules
fragment 227-279
-
fragment consisting of aa 227-279, no severing of microtubules, no binding of microtubules
fragment 227-328
-
fragment consisting of aa 227-328, no severing of microtubules, but binding of microtubules
I406V
-
mutant resembles wild type
K388A
-
mutant, no significant enzymatic activity
N386K
-
disease-associated mutation, no significant enzymatic activity
P45Q
-
leading to an early onset severe form of hereditary spastic paraplegia when present in heterozygosity with a mutant allele
S44L
-
leading to an early onset severe form of hereditary spastic paraplegia when present in heterozygosity with a mutant allele
spastin-DELTAAAA
-
spastin variant without AAA ATPase domain
spastin-DELTACT
-
C-terminal spastin deletion mutant, aa 398-583 deleted
spastin-DELTAexon4
-
spastin variant without exon4
spastin-DELTANT
-
N-terminal spastin deletion mutant, aa 1-300 deleted
A323V
-
mutant dgl1-1, exhibits different response to gibberellin and/or brassinosteroid
D483N
-
mutant dgl1-2, exhibits different response to gibberellin and/or brassinosteroid
DELTA241-408
-
mutant dgl1-3, exhibits different response to gibberellin and/or brassinosteroid
con80
-
cloned cDNA encoding 244 C-terminal P80 amino acids
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
Show AA Sequence (2069 entries)
Please use the Sequence Search for a specific query.