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Literature summary for 5.6.2.4 extracted from
- A novel nuclear DNA helicase with high specific activity from Pisum sativum catalytically translocates in the 3'->5' direction (2003), Eur. J. Biochem., 270, 1735-1745.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| single-stranded DNA | required | Pisum sativum |  |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| actinomycin C1 | - |
Pisum sativum | |
| ammonium sulfate | 45 mM, complete inhibition | Pisum sativum | |
| ATP | the optimum concentration of ATP for DNA helicase activity is 1.0 mM. At 8 mM ATP the DNA unwinding activity of PDH120 is inhibited | Pisum sativum | |
| daunorubicin | - |
Pisum sativum | |
| EDTA | 5 mM, complete inhibition | Pisum sativum | |
| Ethidium bromide | - |
Pisum sativum | |
| KCl | optimum concentration: 250 mM. Completely inhibited at 400 mM | Pisum sativum | |
| M13 dsDNA | 0.03 mM, complete inhibition | Pisum sativum | |
| M13 ssDNA | 0.03 mM, complete inhibition | Pisum sativum | |
| Mg2+ | absolute requirement for divalent cations. Mg2+ at 2.0 mM concentration optimally fulfills this requirement. At 8.0 mM MgCl2 the activity is totally inhibited | Pisum sativum | |
| Nogalamycin | - |
Pisum sativum | |
| potassium phosphate | 100 mM, complete inhibition | Pisum sativum | |
| Trypsin | - |
Pisum sativum |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| nucleus | - |
Pisum sativum | 5634 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| KCl | optimum concentration: 250 mM. Completely inhibited at 400 mM | Pisum sativum | |
| Mg2+ | absolute requirement for divalent cations. Mg2+ at 2.0 mM concentration optimally fulfills this requirement. At 8.0 mM MgCl2 the activity is totally inhibited | Pisum sativum | |
| Mn2+ | 2.0 mM, supports 80% of the activity compared to Mg2+ | Pisum sativum | |
| additional information | Ca2+, Zn2+, Cd2+, Cu2+, Ni2+, Ag2+ and Co2+ are unable to support the activity | Pisum sativum |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 54000 | - |
1 * 54000 + 1 * 66000, SDS-PAGE | Pisum sativum |
| 66000 | - |
1 * 54000 + 1 * 66000, SDS-PAGE | Pisum sativum |
| 120000 | - |
gel filtration, glycerol gradient centrifugation | Pisum sativum |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | Pisum sativum | - |
ADP + phosphate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pisum sativum | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Pisum sativum |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| leaf | - |
Pisum sativum | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
highest specific activity among plant helicases | Pisum sativum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + H2O | - |
Pisum sativum | ADP + phosphate | - |
? | |
| ATP + H2O | the enzyme can unwind 17-bp partial duplex substrates with equal efficiency whether or not they contain a fork. It translocates unidirectionally along the bound strand in the 3' to 5' direction. NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP. The optimum concentration of ATP for DNA helicase activity is 1.0 mM. At 8 mM ATP the DNA unwinding activity of PDH120 is inhibited. No significant difference in the DNA unwinding activity of PDH120 with forked or nonforked substrates. The enzyme fails to unwind synthetic blunt-ended duplex DNA suggesting that PDH120 requires ssDNA adjacent to the duplex as a loading zone | Pisum sativum | ADP + phosphate | - |
? | |
| CTP + H2O | NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP | Pisum sativum | CDP + phosphate | - |
? | |
| dATP + H2O | NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP | Pisum sativum | dADP + phosphate | - |
? | |
| dCTP + H2O | NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP | Pisum sativum | dCDP + phosphate | - |
? | |
| dGTP + H2O | NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP | Pisum sativum | dGDP + phosphate | - |
? | |
| dTTP + H2O | NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP | Pisum sativum | dTDP + phosphate | - |
? | |
| GTP + H2O | NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP | Pisum sativum | GDP + phosphate | - |
? | |
| UTP + H2O | NTPs can support helicase activity in order of decreasing efficiency: ATP, GTP, dCTP, UTP, dTTP, CTP, dATP, dGTP | Pisum sativum | UDP + phosphate | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| heterodimer | 1 * 54000 + 1 * 66000, SDS-PAGE | Pisum sativum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DNA helicase 120 | - |
Pisum sativum |
| PDH120 | - |
Pisum sativum |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 56 | - |
1 min, loss of activity | Pisum sativum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
- |
Pisum sativum |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | 9 | significant unwinding activity is observed in the broad pH range (pH 7.59.0) | Pisum sativum |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.00071 | - |
Nogalamycin | pH 8.0, 37°C | Pisum sativum | |
| 0.004 | - |
daunorubicin | pH 8.0, 37°C | Pisum sativum | |
| 0.0052 | - |
Ethidium bromide | pH 8.0, 37°C | Pisum sativum | |
| 0.0056 | - |
actinomycin C1 | pH 8.0, 37°C | Pisum sativum |
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