Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Oeemig, J.S.; Zhou, D.; Kajander, T.; Wlodawer, A.; Iwai, H.
    NMR and crystal structures of the Pyrococcus horikoshii RadA intein guide a strategy for engineering a highly efficient and promiscuous intein (2012), J. Mol. Biol., 421, 85-99.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.6.4.B7 gene radA, highly efficient cis-splicing of PhoRadA intein, recombinant expression of His-tagged enzyme in Escherichia coli strain ER2566 Pyrococcus horikoshii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.6.4.B7 purified His-tagged wild-type RadA intein and engineered minimized PhoRadA intein mutant, sitting drop vapor diffusion, mixing of 100 nl of protein and 100 nl of precipitant solution at 20°C, X-ray diffraction structure determination and analysis at 1.58-1.75 A resolution, molecular replacement. Comparison between the NMR and crystal structures of PhoRadA intein Pyrococcus horikoshii

Protein Variants

EC Number Protein Variants Comment Organism
3.6.4.B7 additional information structure-based rational design of a functional minimized RadA intein with mutation C1A/T1A (intein/extein) and residues 121-130 removed, the structure of the minimized RadA intein reveals the precise interactions between N-extein and the intein, overview. Effects at the -1 position of N-extein and significant improvement of the splicing efficiency of a less robust splicing variant by eliminating the unfavorable extein-intein interactions observed in the structure. Construction of mutant PhoRad Intein harboring the C1A mutation in the intein and a two-residue C-extein with a mutation of Thr to Ala (T+1A), aimed at prevention of splicing and cleavage Pyrococcus horikoshii

Organism

EC Number Organism UniProt Comment Textmining
3.6.4.B7 Pyrococcus horikoshii O58001 RadA intein; gene radA
-
3.6.4.B7 Pyrococcus horikoshii ATCC 700860 O58001 RadA intein; gene radA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.6.4.B7 recombinant His-tagged wild-type RadA intein and engineered minimized RadA intein from Escherichia coli strain ER2566 by nickel affinity chromatography, dialysis, His-tag cleavage through yeast ubiquitin-like specific protease, and again nickel affinity chromatography to remove the tag, followed by ultrafiltration Pyrococcus horikoshii

Subunits

EC Number Subunits Comment Organism
3.6.4.B7 More primary structures of PhoRadA intein, secondary structure based on the determined NMR model. Comparison between the NMR and crystal structures of PhoRadA intein Pyrococcus horikoshii

Synonyms

EC Number Synonyms Comment Organism
3.6.4.B7 PhoRadA
-
 Pyrococcus horikoshii
3.6.4.B7 RadA
-
 Pyrococcus horikoshii
3.6.4.B7 RadA intein
-
 Pyrococcus horikoshii

General Information

EC Number General Information Comment Organism
3.6.4.B7 evolution cis-splicing of the engineered RadAmin intein is very efficient and indistinguishable from that of PhoRadA intein, suggesting that the removed disordered loop is a mere remnant from the endonuclease domain that was lost during evolution Pyrococcus horikoshii
3.6.4.B7 additional information catalytic site structure analysis, the scissile peptide bond between Met-1 and Ala1 of minimized RadA intein is in the usual transconformation Pyrococcus horikoshii
3.6.4.B7 additional information determination of structures of one of inteins with high splicing efficiency, the RadA intein from Pyrococcus horikoshii (PhoRadA). The solution NMR structure and the crystal structures elucidate the structural basis for its high efficiency, precise interactions between N-extein and the intein, NMR structure determination and structure-function analysis, overview. Comparison between the NMR and crystal structures of PhoRadA intein Pyrococcus horikoshii