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Literature summary for 3.1.21.6 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
Search for more literature for 3.1.21.6

Application

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

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
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
E147A does not exhibit a significant reduction in nicking activity, compared with that of the wild-type Bombyx mori
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
H130A moderately decreased nicking activity Bombyx mori
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
H209A exhibits no detectable cleavage activity on the top or bottom strand of the oligonucleotide substrate Bombyx mori
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
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
K56A moderately decreased nicking activity Bombyx mori
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
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
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
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
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
S43A does not exhibit a significant reduction in nicking activity, compared with that of the wild-type Bombyx mori
S99A does not exhibit a significant reduction in nicking activity, compared with that of the wild-type Bombyx mori
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
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
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]

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

Organism

Organism UniProt Comment Textmining
Bombyx mori
-
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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

Synonyms Comment Organism
R1Bm EN
-
 Bombyx mori
R1Bm endonuclease
-
 Bombyx mori