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Literature summary extracted from

  • Maita, N.; Aoyagi, H.; Osanai, M.; Shirakawa, M.; Fujiwara, H.
    Characterization of the sequence specificity of the R1Bm endonuclease domain by structural and biochemical studies (2007), Nucleic Acids Res., 35, 3918-3927.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
3.1.21.6 additional information important role of residues Y98 and N180 for the sequence recognition of R1Bm EN Bombyx mori

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.21.6 plasmid pR1EN and its mutant derivatives were transformed into Escherichia coli BL21(DE3)/pLysS strain Bombyx mori

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.21.6 by hanging drop vapor diffusion method, to 2.0 A resolution, belongs to space group P321 with unit cell parameters a = b = 141.3 A, c = 37.5 A, alpha = beta = 90°, gamma = 120°, has a beta-hairpin region (residues 176-183) at the edge of the DNA binding surface Bombyx mori

Protein Variants

EC Number Protein Variants Comment Organism
3.1.21.6 D19A relative activity of bottom-strand cleavage is less than half that of top-strand cleavage, whereas activity of top-strand cleavage is more than 80% that of the wild-type Bombyx mori
3.1.21.6 E147A does not exhibit a significant reduction in nicking activity, compared with that of the wild-type Bombyx mori
3.1.21.6 E18A relative activity of bottom-strand cleavage is less than half that of top-strand cleavage, whereas activity of top-strand cleavage is more than 80% that of the wild-type Bombyx mori
3.1.21.6 H130A moderately decreased nicking activity Bombyx mori
3.1.21.6 H140A shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 H209A exhibits no detectable cleavage activity on the top or bottom strand of the oligonucleotide substrate Bombyx mori
3.1.21.6 H54A relative activity of bottom-strand cleavage is less than half that of top-strand cleavage, whereas activity of top-strand cleavage is more than 80% that of the wild-type Bombyx mori
3.1.21.6 H78A shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 K56A moderately decreased nicking activity Bombyx mori
3.1.21.6 N180A altered cleavage patterns, shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 Q97A shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 R139A shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 R144A shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 S206A relative activity of bottom-strand cleavage is less than half that of top-strand cleavage, whereas activity of top-strand cleavage is more than 80% that of the wild-type Bombyx mori
3.1.21.6 S43A does not exhibit a significant reduction in nicking activity, compared with that of the wild-type Bombyx mori
3.1.21.6 S99A does not exhibit a significant reduction in nicking activity, compared with that of the wild-type Bombyx mori
3.1.21.6 T178A shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 Y42A shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori
3.1.21.6 Y98A altered cleavage patterns, shows less than 20% of the wild-type activity for the cleavage of both strands, amounts of cleavage products are less than 1% of the total substrates Bombyx mori

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.21.6 22000
-
 gel filtration Bombyx mori

Organism

EC Number Organism UniProt Comment Textmining
3.1.21.6 Bombyx mori
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.21.6 by nickel chelation chromatography and by gel filtration Bombyx mori

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.21.6 28S rDNA + H2O R1Bm EN cleaves both strands of the 28S rDNA sequence at the target site, 5'-A(G/C)(A/T)!(A/G)T-3' is the consensus sequence, thus cleavage site Bombyx mori ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.21.6 R1Bm EN
-
 Bombyx mori
3.1.21.6 R1Bm endonuclease
-
 Bombyx mori