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Information on EC 3.6.5.5 - dynamin GTPase

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EC Tree
IUBMB Comments
An enzyme with a molecular mass of about 100 kDa that is involved in endocytosis and is instrumental in pinching off membrane vesicles.
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This record set is specific for:
UNIPROT: O00429
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Word Map
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
Synonyms
dynamin, dynamin-related protein 1, dynamin 2, d100, dynamin-2, dynamin 1, dynamin i, optic atrophy 1, dynamin-1, gtpase dynamin, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Drp1-short
isozyme 3
dynamin GTPase
-
dynamin-1
-
dynamin-1-like GTPase
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dynamin-related protein 1
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dynamin-related protein 1 isoform 3
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fission dynamin
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hDrp1 isoform 3
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B-dynamin
-
-
-
-
D100
-
-
-
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Dynamin BREDNM19
-
-
-
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Dynamin UDNM
-
-
-
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Dynamin, brain
-
-
-
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Dynamin, testicular
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-
-
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GTP phosphohydrolase
-
-
-
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GTPase
-
-
-
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guanine triphosphatase
-
-
-
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guanosine 5'-triphosphatase
-
-
-
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guanosine triphosphatase
-
-
-
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phosphatase, guanosine tri-
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-
-
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ribosomal GTPase
-
-
-
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Shibire protein
-
-
-
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T-dynamin
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
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-
-
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SYSTEMATIC NAME
IUBMB Comments
GTP phosphohydrolase (vesicle-releasing)
An enzyme with a molecular mass of about 100 kDa that is involved in endocytosis and is instrumental in pinching off membrane vesicles.
CAS REGISTRY NUMBER
COMMENTARY hide
9059-32-9
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
GTP + H2O
GDP + phosphate
show the reaction diagram
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
GTP + H2O
GDP + phosphate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
actin
cardiolipin
ionomycin
a calcium ionophore, causes rapid mitochondrial accumulation of actin filaments followed by Drp1 accumulation at the fission site, and increases fission rate
Mff protein
mitocjondrial protein Mff alone causes only a slight increase in Drp1 GTPase activity at the concentrations tested, while the combination of Mff and actin filaments causes a substantial increase in Drp1 activity, far beyond the additive effects of either Mff or actin alone
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Mff-proteoliposome
lipid-stimulated GTPase activity
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mitochondrial fission factor
Mff, stimulates Drp1 independently of, but synergistically with cardiolipin
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additional information
interchangeable adaptors regulate mitochondrial dynamin assembly for membrane scission
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.4
assay at
7.5
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23
GTPase and polymerization assay at
30
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
Drp1 is a cytosolic protein that translocates to the outer mitochondrial membrane
Manually annotated by BRENDA team
Drp1 is a cytosolic protein that translocates to the outer mitochondrial membrane
Manually annotated by BRENDA team
additional information
actin filaments target the oligomeric maturation of the dynamin GTPase Drp1 to mitochondrial fission sites. Cytosolic Drp1 is recruited directly to fission sites immediately prior to fission. Progressive maturation of Drp1 oligomers on mitochondria through incorporation of smaller mitochondrially-bound Drp1 units. Maturation of a stable Drp1 oligomer does not forcibly lead to fission. Drp1 oligomers also translocate directionally along mitochondria. Drp1 is a cytosolic protein that translocates to the outer mitochondrial membrane
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme is a member of the dynamin superfamily that comprises a family of conserved GTPases. Most dynamin superfamily members contain five conserved GTP-binding motifs (G1-5), similar to small Ras-like GTPases
malfunction
metabolism
the enzyme is a key regulator of mitochondrial fission
physiological function
additional information
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
DNM1L_HUMAN
736
0
81877
Swiss-Prot
other Location (Reliability: 4)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
169624
recombinant enzyme, analytical equilibrium sedimentation analysis
40923
x * 40923, nucleotide-free GTPase-GTPase effector domain fusion protein of Drp1, mass spectrometry
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 40923, nucleotide-free GTPase-GTPase effector domain fusion protein of Drp1, mass spectrometry
dimer
2 * 78000-85000, recombinant enzyme, SDS-PAGE
oligomer
in solution, Drp1 exists in a number of oligomeric states, including dimers, tetramers, and higher-order oligomers. Actin filaments can organize Drp1 into a productive oligomer
additional information
Drp1 isoforms exhibit differential oligomerization propensities in solution and distinct helical geometry on cardiolipin-containing membranes, analysis of Drp1-short, Drp1-A-only, and Drp1-B-only by gel filtration
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
actin filaments target the oligomeric maturation of the dynamin GTPase Drp1 to mitochondrial fission sites
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant dynamin-related protein 1 GTPase-GTPase effector domain fusion protein, sitting drop vapour diffusion method, mixing of 0.0004 ml of protein solution and reservoir solution, the latter containing 0.2 M lithium sulfate, 0.1 M Bis-Tris, pH 5.5, 25% v/v PEG 3350 or 0.1 M sodium citrate, pH 5.5, 20% PEG 3000, 20°C, X-ray diffraction structure determination and analysis at 2.67 A resolution
purified wild-type and mutants of DNM1L isoform 2, mixing of 0.001 ml of 1.2 mg/ml protein with 0.001 ml of reservoir solution containing 0.1 M sodium citrate pH 5, 27.5% PEG 3000, equilibration against 0.4 ml of reservoir solution, 3-5 days, for co-crystallization of the enzyme with a non-hydrolyzable GTP analogue the protein is incubated with 1 mM GMP-PNP and 4 mM MgCl2, and then purified by gel filtration, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement using human dynamin-1 structure, PDB ID 3SNH
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D146A
site-directed mutagenesis
D190A
site-directed mutagenesis
D218A
site-directed mutagenesis
E81A
site-directed mutagenesis
E81A/E82A
site-directed mutagenesis
G149A
site-directed mutagenesis
K216A
site-directed mutagenesis
N246A
site-directed mutagenesis
Q34A
site-directed mutagenesis
S35A
site-directed mutagenesis
S39A
site-directed mutagenesis
S40A
site-directed mutagenesis
T59A
site-directed mutagenesis
additional information
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant N-terminally Strep-tagged Drp1 from Escherichia coli strain BL21 Star (DE3) by affinity chromatography, followed by tag cleavage by HRV3C protease, and gel filtration
recombinant N-terminally Strep-tagged Drp1 from Saccharomyces cerevisiae strain JSY9612 by affinity chromatography, followed by tag cleavage by PreScission protease, and dialysis
recombinant nucleotide-free GTPase-GTPase effector domain fusion protein of human Drp1, and wild-type and mutant full-length enzymes from Escherichia coli strain BL21(DE3)
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene DNM1, recombinant expression of Drp1 containing an N-terminal PreScission protease cleavage site and a FLAG-One-Strep-tag (IBA) from plasmid pRS415-DNM1 in Saccharomyces cerevisiae strain JSY9612 lacking all fission proteins (MATa, can1, ade2, trp1, ura3, his3, leu2, pep4::HIS3, prb1::LEU2, bar1::HISG, lys2::GAL1/10-GAL4), coexpression with Saccharomyces cerevisiae mitochondrial fission 1 protein, Fis1 (UniProt ID P40515)
gene DNMTL1, recombinant expression of the splice variants of Drp1 in Escherichia coli strain BL21 Star (DE3)
recombinant expression of C-terminally His6-tagged wild-type and mutants of DNM1L isoform 2
recombinant expression of N-terminally Strep-tagged Drp1 with an HRV3C protease site from pET16b vector in Escherichia coli strain BL21 Star (DE3), stable recombinant expression of GFP-tagged DrpI in U2OS cells with partial shRNA suppression of endogenous Drp1 (gDrp-U2OS), GFP-Drp1 functionally compensates for endogenous Drp1 in these cells
recombinant expression of nucleotide-free GTPase-GTPase effector domain fusion protein of enzyme Drp1, and wild-type and mutant full-length enzymes in Escherichia coli strain BL21(DE3)
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Klinglmayr, E.; Wenger, J.; Mayr, S.; Bossy-Wetzel, E.; Puehringer, S.
Purification, crystallization and X-ray diffraction analysis of human dynamin-related protein 1 GTPase-GED fusion protein
Acta Crystallogr. Sect. F
68
1217-1221
2012
Homo sapiens (O00429), Homo sapiens
Manually annotated by BRENDA team
Wenger, J.; Klinglmayr, E.; Froehlich, C.; Eibl, C.; Gimeno, A.; Hessenberger, M.; Puehringer, S.; Daumke, O.; Goettig, P.
Functional mapping of human dynamin-1-like GTPase domain based on x-ray structure analyses
PLoS ONE
8
e71835
2013
Homo sapiens (O00429), Homo sapiens
Manually annotated by BRENDA team
Ji, W.K.; Hatch, A.L.; Merrill, R.A.; Strack, S.; Higgs, H.N.
Actin filaments target the oligomeric maturation of the dynamin GTPase Drp1 to mitochondrial fission sites
eLife
4
e11553
2015
Homo sapiens (O00429)
Manually annotated by BRENDA team
Macdonald, P.J.; Francy, C.A.; Stepanyants, N.; Lehman, L.; Baglio, A.; Mears, J.A.; Qi, X.; Ramachandran, R.
Distinct splice variants of dynamin-related protein 1 differentially utilize mitochondrial fission factor as an effector of cooperative GTPase activity
J. Biol. Chem.
291
493-507
2016
Homo sapiens (O00429), Rattus norvegicus (O35303)
Manually annotated by BRENDA team
Koirala, S.; Guo, Q.; Kalia, R.; Bui, H.T.; Eckert, D.M.; Frost, A.; Shaw, J.M.
Interchangeable adaptors regulate mitochondrial dynamin assembly for membrane scission
Proc. Natl. Acad. Sci. USA
110
E1342-E1351
2013
Homo sapiens (O00429), Saccharomyces cerevisiae (P54861), Saccharomyces cerevisiae
Manually annotated by BRENDA team