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Literature summary for 3.1.30.2 extracted from

  • Anisimova, V.E.; Shcheglov, A.S.; Bogdanova, E.A.; Rebrikov, D.V.; Nekrasov, A.N.; Barsova, E.V.; Shagin, D.A.; Lukyanov, S.A.
    Is crab duplex-specific nuclease a member of the Serratia family of non-specific nucleases? (2008), Gene, 418, 41-48.
    View publication on PubMed

Cloned(Commentary)

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

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/ Products (Substrates)

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 ?
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?

Organism

Organism UniProt Comment Textmining
Paralithodes camtschaticus Q8I9M9
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Purification (Commentary)

Purification (Comment) Organism
Par_DSN-t1 is insoluble and inactive after puriciation and renaturation Paralithodes camtschaticus

Renatured (Commentary)

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 [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
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G133V mutant, dsDNA, not detectable (inactive protein) Paralithodes camtschaticus
additional information
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G139V mutant, dsDNA, not detectable (inactive protein) Paralithodes camtschaticus
additional information
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H168A mutant, dsDNA, not detectable (inactive protein) Paralithodes camtschaticus
additional information
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H171A mutant, dsDNA, not detectable (inactive protein) Paralithodes camtschaticus
additional information
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H237A mutant, dsDNA, not detectable Paralithodes camtschaticus
additional information
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K235A mutant, dsDNA, 110 Kunitz units Paralithodes camtschaticus
additional information
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K235R mutant, dsDNA, 5980 Kunitz units Paralithodes camtschaticus
additional information
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Par_DSN-t1 lacks DNAse activity Paralithodes camtschaticus
additional information
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Par_DSN-t2 lacks DNase activity Paralithodes camtschaticus
additional information
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Par_DSN-t3 lacks DNAse activity Paralithodes camtschaticus
additional information
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Par_DSN-t4 has low specific activity (6 Kunitz-units compared to 6070 Kunitz units for intact Par_DSN), dsDNA Paralithodes camtschaticus
additional information
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R121/122A mutant, dsDNA, not detectable (inactive protein) Paralithodes camtschaticus
additional information
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R121A mutant, dsDNA, 5960 Kunitz units Paralithodes camtschaticus
additional information
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R121K mutant, dsDNA, 5990 Kunitz units Paralithodes camtschaticus
additional information
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R122A mutant, dsDNA, 6030 Kunitz units Paralithodes camtschaticus
additional information
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R122K mutant, dsDNA, 6050 Kunitz units Paralithodes camtschaticus
additional information
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R184A mutant, dsDNA, 1810 Kunitz units Paralithodes camtschaticus
additional information
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R184K mutant, dsDNA, 6100 Kunitz units Paralithodes camtschaticus
additional information
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wild-type, dsDNA, 6070 Kunitz units Paralithodes camtschaticus

Substrates and Products (Substrate)

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 ?
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?

Synonyms

Synonyms Comment Organism
Kamchatka crab duplex-specific nuclease
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Paralithodes camtschaticus
Par_DSN novel subfamily of bba-Me-finger Paralithodes camtschaticus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
Par_DSN-t4 Paralithodes camtschaticus
60
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Par_DSN Paralithodes camtschaticus

Temperature Stability [°C]

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