Cloned (Comment) | Organism |
---|---|
Par_DSN-t1: residues 192-379 of Par_DSN precursor (GenBank ID AAN86143) linked with a bacterial signal peptide and 6×His tag (sequence corresponds to the NUC domain). Par_DSN-t2: residues 149-406, corresponds to the NUC domain identified by multiple sequence alignment, is sensitive to proteinase K and is completely cleaved by this protease. Par_DSN-t3: residues 162-406, additionaly all unpaired Cys residues are deleted, Par_DSN-t3 is soluble, sensitive to proteinase K. Par_DSN-t4: residues 114-406, Par_DSN-t4 is degraded by proteinase K | Paralithodes camtschaticus |
Protein Variants | Comment | Organism |
---|---|---|
G133V | yield after proteinase K treatment 0% | Paralithodes camtschaticus |
G139V | yield after proteinase K treatment 0% | Paralithodes camtschaticus |
H168A | yield after proteinase K treatment 0% | Paralithodes camtschaticus |
H171A | yield after proteinase K treatment 0% | Paralithodes camtschaticus |
H237A | yield after proteinase K treatment 20% | Paralithodes camtschaticus |
K235A | yield after proteinase K treatment 19% | Paralithodes camtschaticus |
K235R | yield after proteinase K treatment 21% | Paralithodes camtschaticus |
R121/122A | yield after proteinase K treatment 4% | Paralithodes camtschaticus |
R121A | for specific activity measurement | Paralithodes camtschaticus |
R121K | yield after proteinase K treatment 19% | Paralithodes camtschaticus |
R122A | for specific activity measurement | Paralithodes camtschaticus |
R122K | yield after proteinase K treatment 20% | Paralithodes camtschaticus |
R184A | yield after proteinase K treatment 22% | Paralithodes camtschaticus |
R184K | yield after proteinase K treatment 18% | Paralithodes camtschaticus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
DNA + H2O | Paralithodes camtschaticus | dsDNA to ssDNA preference ratio for Par_DSN-t4 is equal to 10, corresponding ratio for intact Par_DSN is equal to 1000 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paralithodes camtschaticus | Q8I9M9 | - |
- |
Purification (Comment) | Organism |
---|---|
Par_DSN-t1 is insoluble and inactive after puriciation and renaturation | Paralithodes camtschaticus |
Renatured (Comment) | Organism |
---|---|
Par_DSN-t1 is insoluble and inactive after purification and renaturation. Par_DSN-t2, Par_DSN-t3, Par_DSN-t4 could be renaturated. | Paralithodes camtschaticus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
G133V mutant, dsDNA, not detectable (inactive protein) | Paralithodes camtschaticus |
additional information | - |
G139V mutant, dsDNA, not detectable (inactive protein) | Paralithodes camtschaticus |
additional information | - |
H168A mutant, dsDNA, not detectable (inactive protein) | Paralithodes camtschaticus |
additional information | - |
H171A mutant, dsDNA, not detectable (inactive protein) | Paralithodes camtschaticus |
additional information | - |
H237A mutant, dsDNA, not detectable | Paralithodes camtschaticus |
additional information | - |
K235A mutant, dsDNA, 110 Kunitz units | Paralithodes camtschaticus |
additional information | - |
K235R mutant, dsDNA, 5980 Kunitz units | Paralithodes camtschaticus |
additional information | - |
Par_DSN-t1 lacks DNAse activity | Paralithodes camtschaticus |
additional information | - |
Par_DSN-t2 lacks DNase activity | Paralithodes camtschaticus |
additional information | - |
Par_DSN-t3 lacks DNAse activity | Paralithodes camtschaticus |
additional information | - |
Par_DSN-t4 has low specific activity (6 Kunitz-units compared to 6070 Kunitz units for intact Par_DSN), dsDNA | Paralithodes camtschaticus |
additional information | - |
R121/122A mutant, dsDNA, not detectable (inactive protein) | Paralithodes camtschaticus |
additional information | - |
R121A mutant, dsDNA, 5960 Kunitz units | Paralithodes camtschaticus |
additional information | - |
R121K mutant, dsDNA, 5990 Kunitz units | Paralithodes camtschaticus |
additional information | - |
R122A mutant, dsDNA, 6030 Kunitz units | Paralithodes camtschaticus |
additional information | - |
R122K mutant, dsDNA, 6050 Kunitz units | Paralithodes camtschaticus |
additional information | - |
R184A mutant, dsDNA, 1810 Kunitz units | Paralithodes camtschaticus |
additional information | - |
R184K mutant, dsDNA, 6100 Kunitz units | Paralithodes camtschaticus |
additional information | - |
wild-type, dsDNA, 6070 Kunitz units | Paralithodes camtschaticus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
DNA + H2O | dsDNA to ssDNA preference ratio for Par_DSN-t4 is equal to 10, corresponding ratio for intact Par_DSN is equal to 1000 | Paralithodes camtschaticus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Kamchatka crab duplex-specific nuclease | - |
Paralithodes camtschaticus |
Par_DSN | novel subfamily of bba-Me-finger | Paralithodes camtschaticus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
Par_DSN-t4 | Paralithodes camtschaticus |
60 | - |
Par_DSN | Paralithodes camtschaticus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
activity of Par_DSN-t4 completely disappears after incubation for 20 min, full-length Par_DSN retains 25% activity | Paralithodes camtschaticus |