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Literature summary extracted from
- The C-terminus of kinesin-14 Ncd is a crucial component of the force generating mechanism (2012), FEBS Lett., 586, 854-858.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.6.1.4 | additional information | microtubule-stimulated ATPase | Drosophila melanogaster |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.6.1.4 | His tagged truncated proteins (e.g. wild type, residue 250-700) expressed in Escherichia coli | Drosophila melanogaster |
| 5.6.1.4 | recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli | Drosophila melanogaster |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.6.1.4 | M672N | site-directed mutagenesis, the mutation has a negligible effect on microtubule binding in the absence of nucleotides although substantially changed the Kd(MT) in the presence of AMPPNP | Drosophila melanogaster |
| 5.6.1.4 | additional information | truncated versions: residue 250-700 (wild type), residue 346-700 (motor domain 700), residue 250-670 (Ncd670), residue 346-700 (motor domain 670), residue 250-679 (Ncd679), residue 250-670 + GGSGGSGGS + 680-700 (Ncd700SG, residue 671-679 replaced by sequence GGSGGSGGS), residue 250-670 with M672N exchange (NcdM672N) | Drosophila melanogaster |
| 5.6.1.4 | additional information | production of four Ncd mutants in which the C-terminus is altered or deleted, analysis of mutant affinity to the microtubule, steady-state ATPase and gliding velocity in multiple motor assays: the mutations have a dramatic effect on all three parameters measured, suggesting that the C-terminal residues of Ncd play an important role in modulating the interaction of the motor with the microtubule. In the first of the mutants, Ncd670, the entire C-terminal segment is deleted. In mutant Ncd679, the basic fragment spanning aa 671679 is left intact whereas the last 21 aa are deleted. In mutant Ncd700SG the basic fragment is replaced by the sequence GGSGGSGGS, which is uncharged and flexible. In the fourth construct NcdM672N, the hydrophobic Met672 is substituted by a small and polar Asn | Drosophila melanogaster |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.6.1.4 | 0.00014 | - |
ATP | mutants Ncd670 and Ncd700SG, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster |  |
| 5.6.1.4 | 0.00022 | - |
ATP | wild-type Ncd, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
| 5.6.1.4 | 0.00026 | - |
ATP | mutant Ncd679, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
| 5.6.1.4 | 0.00061 | - |
ATP | mutant NcdM672N, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 5.6.1.4 | microtubule | - |
Drosophila melanogaster | 5874 | - |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.6.1.4 | Mg2+ | required | Drosophila melanogaster | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.6.1.4 | ATP + H2O + a kinesin associated with a microtubule at position n | Drosophila melanogaster | - |
ADP + phosphate + a kinesin associated with a microtubule at position n-1 (toward the minus end) | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.6.1.4 | Drosophila melanogaster | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.6.1.4 | immobilized metal ion affinity chromatography (Ni2+) | Drosophila melanogaster |
| 5.6.1.4 | recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli | Drosophila melanogaster |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.6.1.4 | ATP + H2O + a kinesin associated with a microtubule at position n | - |
Drosophila melanogaster | ADP + phosphate + a kinesin associated with a microtubule at position n-1 (toward the minus end) | - |
? | |
| 5.6.1.4 | additional information | measurement of Ncd affinity to microtubules, microtubule-stimulated ATPase, and motility of wild-type and mutant enzymes | Drosophila melanogaster | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 5.6.1.4 | dimer | - |
Drosophila melanogaster |
| 5.6.1.4 | homodimer | - |
Drosophila melanogaster |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.6.1.4 | kinesin-14 | - |
Drosophila melanogaster |
| 5.6.1.4 | kinesin-14 Ncd | - |
Drosophila melanogaster |
| 5.6.1.4 | ncd | - |
Drosophila melanogaster |
| 5.6.1.4 | non-claret disjunctional | - |
Drosophila melanogaster |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.6.1.4 | 25 | - |
assay at | Drosophila melanogaster |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.6.1.4 | 0.62 | - |
ATP | Ncd700SG (residue 250-700, residue 671-679 replaced by sequence GGSGGSGGS), microtuble stimulated, Vmax/2 at 0.00014 mM microtubles, pH 7.2, 25°C | Drosophila melanogaster | |
| 5.6.1.4 | 0.62 | - |
ATP | mutant Ncd700SG, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
| 5.6.1.4 | 0.83 | - |
ATP | Ncd670 (residue 250-670), microtuble stimulated, Vmax/2 at 0.00014 mM microtubles, pH 7.2, 25°C | Drosophila melanogaster | |
| 5.6.1.4 | 0.83 | - |
ATP | mutant Ncd670, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
| 5.6.1.4 | 2.49 | - |
ATP | NcdM672N (residue 250-679, M672N mutation), microtuble stimulated, Vmax/2 at 0.00061 mM microtubles, pH 7.2, 25°C | Drosophila melanogaster | |
| 5.6.1.4 | 2.49 | - |
ATP | mutant NcdM672N, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
| 5.6.1.4 | 2.58 | - |
ATP | NcdWT (residue 250-700), microtuble stimulated, Vmax/2 at 0.00022 mM microtubles, pH 7.2, 25°C | Drosophila melanogaster | |
| 5.6.1.4 | 2.58 | - |
ATP | wild-type Ncd, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
| 5.6.1.4 | 3.58 | - |
ATP | Ncd679 (residue 250-679), microtuble stimulated, Vmax/2 at 0.00026 mM microtubles, pH 7.2, 25°C | Drosophila melanogaster | |
| 5.6.1.4 | 3.58 | - |
ATP | mutant Ncd679, pH 7.2, 25°C, microtubule-stimulated ATPase activity | Drosophila melanogaster | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.6.1.4 | 7.2 | - |
assay at | Drosophila melanogaster |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 5.6.1.4 | evolution | Ncd is a member of kinesin superfamily, kinesin-14 subfamily motors that uses the power stroke, a lever-like pivoting action of a long and stiff element, to exert force and generate movement | Drosophila melanogaster |
| 5.6.1.4 | additional information | the C-terminal segment of the kinesin-14 subfamily is formed by residues that extend beyond the a6 helix | Drosophila melanogaster |
| 5.6.1.4 | physiological function | Ncd is a member of kinesin-14 subfamily motors that uses the power stroke, a lever-like pivoting action of a long and stiff element, to exert force and generate movement, the Ncd C-terminus is crucial in this process modulating the interaction of the motor with the microtubule. Ncd is a retrograde motor that participates in spindle assembly by generating microtubule-microtubule sliding in an orientation dependent manner | Drosophila melanogaster |
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