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Literature summary extracted from

  • Nagasawa, Y.; Fujii, K.; Yoshikawa, T.; Kobayashi, Y.; Kondo, T.
    Pokeweed antiviral protein region Gly209-Lys225 is critical for RNA N-glycosidase activity of the prokaryotic ribosome (2008), Phytochemistry, 69, 1653-1660.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.2.22 plasmid pMAL-c2 to amplify pokeweed antiviral protein and karasurin-A by PCR, expression in Escherichia coli XL1-Blue cells with expression plasmid pGEX-6P-1 Phytolacca americana
3.2.2.22 plasmid pMAL-c2 to amplify pokeweed antiviral protein and karasurin-A by PCR, expression in Escherichia coli XL1-Blue cells with expression plasmid pGEX-6P-1 Trichosanthes kirilowii

Protein Variants

EC Number Protein Variants Comment Organism
3.2.2.22 additional information chimera proteins of 3 domains of pokeweed antiviral protein (PAP) and karasurin-A (N-terminal 1-74, central 75-177, C-terminal 178-262), C-terminal end determines determines specificity to eukaryotic or eukaryotic and prokaryotic ribosomes, PAP-region 209-225 is required for activity towards prokaryotic ribosomes Phytolacca americana
3.2.2.22 additional information chimera proteins of 3 domains of pokeweed antiviral protein and karasurin-A (N-terminal 1-74, central 75-177, C-terminal 178-262), C-terminal end determines determines specificity to eukaryotic or eukaryotic and prokaryotic ribosomes, PAP-region 209-225 is required for activity towards prokaryotic ribosomes Trichosanthes kirilowii

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.2.22 additional information Trichosanthes kirilowii karasurin-A shows RNA N-glycosidase activity only towards eukaryotic ribosomes while pokeweed antiviral protein is active towards eukaryotic and prokaryotic ribosomes, removal of specific adenine from alpha-sarcin/ricin loop of the large rRNA, substrate specificity (eukaryotic/prokaryotic) determined by C-terminus residues 209-225 ?
-
 ?
3.2.2.22 additional information Phytolacca americana PAP shows RNA N-glycosidase activity towards eukaryotic and prokaryotic ribosomes, karasurin-A only towards eukaryotic ribosomes, removal of specific adenine from alpha-sarcin/ricin loop of the large rRNA, substrate specificity (eukaryotic/prokaryotic) determined by C-terminus residues 209-225 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.2.22 Phytolacca americana
-
 pokeweed
-
3.2.2.22 Trichosanthes kirilowii P24478 var. japonica
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.2.22 cell centrifugation, washing, lysozyme application, sonication, centrifugation, supernatants applied to GSTrap FF column, elution with sonication buffer and 10 mM reduced glutathione, desalted, concentrated and glutathione S-transferase (GST) cleaved off with PreScission Protease Phytolacca americana
3.2.2.22 cell centrifugation, washing, lysozyme application, sonication, centrifugation, supernatants applied to GSTrap FF column, elution with sonication buffer and 10 mM reduced glutathione, desalted, concentrated and glutathione S-transferase (GST) cleaved off with PreScission Protease Trichosanthes kirilowii

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.2.22 leaf Phytolacca americana Phytolacca americana
-
3.2.2.22 root
-
 Trichosanthes kirilowii
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.2.22 additional information karasurin-A shows RNA N-glycosidase activity only towards eukaryotic ribosomes while pokeweed antiviral protein is active towards eukaryotic and prokaryotic ribosomes, removal of specific adenine from alpha-sarcin/ricin loop of the large rRNA, substrate specificity (eukaryotic/prokaryotic) determined by C-terminus residues 209-225 Trichosanthes kirilowii ?
-
 ?
3.2.2.22 additional information PAP shows RNA N-glycosidase activity towards eukaryotic and prokaryotic ribosomes, karasurin-A only towards eukaryotic ribosomes, removal of specific adenine from alpha-sarcin/ricin loop of the large rRNA, substrate specificity (eukaryotic/prokaryotic) determined by C-terminus residues 209-225 Phytolacca americana ?
-
 ?
3.2.2.22 RNA + H2O ribosomes from rat liver (eukaryotic) and Escherichia coli JM109 (prokaryotic), 37°C, 25 mM Tris-HCl buffer, pH 7.6 with 50 mM KCl and 5 mM MgCl2 Phytolacca americana ?
-
 ?
3.2.2.22 RNA + H2O ribosomes from rat liver (eukaryotic) and Escherichia coli JM109 (prokaryotic), 37°C, 25 mM Tris-HCl buffer, pH 7.6 with 50 mM KCl and 5 mM MgCl2 Trichosanthes kirilowii ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.2.22 karasurin-A KRN recombinantly combined with pokeweed antiviral protein (PAP) Trichosanthes kirilowii
3.2.2.22 PAP recombinantly combined with karasurin-A (KRN) of Trichosanthes kirilowii Phytolacca americana
3.2.2.22 pokeweed antiviral protein
-
 Phytolacca americana