Cloned (Comment) | Organism |
---|---|
recombinant expression of C-terminally His6-tagged wild-type and mutants of DNM1L isoform 2 | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
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 | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D146A | site-directed mutagenesis | Homo sapiens |
D190A | site-directed mutagenesis | Homo sapiens |
D218A | site-directed mutagenesis | Homo sapiens |
E81A | site-directed mutagenesis | Homo sapiens |
E81A/E82A | site-directed mutagenesis | Homo sapiens |
G149A | site-directed mutagenesis | Homo sapiens |
K216A | site-directed mutagenesis | Homo sapiens |
K38A | site-directed mutagenesis | Homo sapiens |
N246A | site-directed mutagenesis | Homo sapiens |
Q34A | site-directed mutagenesis | Homo sapiens |
S35A | site-directed mutagenesis | Homo sapiens |
S39A | site-directed mutagenesis | Homo sapiens |
S40A | site-directed mutagenesis | Homo sapiens |
T59A | site-directed mutagenesis | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required for activity | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + H2O | Homo sapiens | - |
GDP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | O00429 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + H2O | - |
Homo sapiens | GDP + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DNM1L | - |
Homo sapiens |
dynamin-1-like GTPase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | 7.4 | assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
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 | Homo sapiens |
malfunction | mutated Glu81 and Glu82 in the unique 16-residue insertion of the enzyme influence the activity significantly. Mutations of Gln34, Ser35, and Asp190 in the predicted assembly interface interfer with dimerization of the GTPase domain induced by a transition state analogue and lead to a loss of the lipid-stimulated GTPase activity | Homo sapiens |
additional information | the GTPase domain of the enzyme is structurally related to that of dynamin and binds the nucleotide 5'-guanylyl-imidodiphosphate (GMP-PNP) via five highly conserved motifs, whereas the BSE folds into a pocket at the opposite side. Residues essential for the GTPase reaction are Lys38, Ser39 and Ser40 in the phosphate binding loop, Thr59 from switch I, Asp146 and Gly149 from switch II, Lys216 and Asp218 in the G4 element, as well as Asn246 in the G5 element. Conformational changes upon GTP-binding in the active site, and overall structure of the DNM1L GTPase-GED fusion protein, overview | Homo sapiens |
physiological function | human dynamin-1-like protein is a GTP-driven molecular machine that segregates mitochondria and peroxisomes, catalytic mechanism involving dimerization of their GTPase domains, overview | Homo sapiens |