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Literature summary extracted from

  • Okuda, M.; Shiba, T.; Inaoka, D.K.; Kita, K.; Kurisu, G.; Mineki, S.; Harada, S.; Watanabe, Y.; Yoshinari, S.
    A conserved lysine residue in the crenarchaea-specific loop is important for the crenarchaeal splicing endonuclease activity (2011), J. Mol. Biol., 405, 92-104.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.6.1.16 expression of His6-tagged wild-type and mutant EndAs in Escherichi coli strain Rosetta 2(DE3) Aeropyrum pernix

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.6.1.16 purified recombinant wild-type and mutant EndAs, method screening, sitting drop vapor diffusion technique, mixing of 0.001 ml protein solution with 0.001 ml reservoir solution containing 0.2 M NaCl, 0.1 M phosphate-citrate, pH 4.2, and 10% w/v PEG 3000, equilibration over 0.1 ml reservoir solution, 22°C, X-ray diffraction structure determmination and analysis at 1.7-2.3 A resolution, molecular replacement Aeropyrum pernix

Protein Variants

EC Number Protein Variants Comment Organism
4.6.1.16 D49A site-directed mutagenesis, the mutation does not affect the enzyme activity Aeropyrum pernix
4.6.1.16 E43A site-directed mutagenesis, the mutation does not affect the enzyme activity Aeropyrum pernix
4.6.1.16 E51A site-directed mutagenesis, the mutation does not affect the enzyme activity Aeropyrum pernix
4.6.1.16 F50A site-directed mutagenesis, the mutant shows reduced enzyme activity compared to the wild-type enzyme Aeropyrum pernix
4.6.1.16 H133A site-directed mutagenesis, a catalytic site mutant, crystal structure determination Aeropyrum pernix
4.6.1.16 K44A site-directed mutagenesis, the mutant shows almost no enzyme activity Aeropyrum pernix
4.6.1.16 P45A site-directed mutagenesis, the mutant shows reduced enzyme activity compared to the wild-type enzyme Aeropyrum pernix
4.6.1.16 R46A site-directed mutagenesis, the mutant shows reduced enzyme activity compared to the wild-type enzyme Aeropyrum pernix

Organism

EC Number Organism UniProt Comment Textmining
4.6.1.16 Aeropyrum pernix Q9YBF1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.6.1.16 recombinant wild-type and mutant EndAs from Escherichia coli strain Rosetta 2(DE3) by heat treatment at 70°C for 30 min, followed by metal affinity chromatography and gel filtration Aeropyrum pernix

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.6.1.16 additional information Sulfolobus tokodaii tRNATrp precursor as a substrate Aeropyrum pernix ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.6.1.16 EndA
-
 Aeropyrum pernix
4.6.1.16 splicing endonuclease
-
 Aeropyrum pernix

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.6.1.16 70
-
 assay at Aeropyrum pernix

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.6.1.16 8
-
 assay at Aeropyrum pernix

General Information

EC Number General Information Comment Organism
4.6.1.16 evolution the crenarchaeal heterotetrameric EndAs can be further classified into two subfamilies based on the size of the structural subunit. Subfamily A possesses a structural subunit similar in size to the catalytic subunit, whereas subfamily B possesses a structural subunit significantly smaller than the catalytic subunit Aeropyrum pernix
4.6.1.16 additional information EndA from Aeropyrum pernix also possesses an extra loop region that is characteristic of crenarchaeal EndAs, the conserved lysine residue Lys44 in the loop is important for endonuclease activity, substrate docking modeling, overview Aeropyrum pernix