3.2.2.22: rRNA N-glycosylase

This is an abbreviated version, for detailed information about rRNA N-glycosylase, go to the full flat file.

Reaction

hydrolysis of the N-glycosylic bond at A-4324 in 28S rRNA from rat ribosomes =

Synonyms

(ribosome-inactivating protein)-like protein, abelesculin, ABRaA, abrin, abrin-a A chain, Abrus precatorius agglutinin, Agglutinin, alpha-MC, alpha-momorcharin, alpha-pisavin, aralin, B-32, balsamin, BBAP1, beta-momorcharin, beta-pisavin, Bougainvillea xbuttiana antiviral protein 1, bouganin, camphorin, CAP30, cinnamomin, Cochinin B, cytotoxic ribosome-inactivating lectin, depurinating rRNA N-glycosidase, dianthin 30, dianthin 32, diphtheria toxin, ebulin b, ebulitin alpha, ebulitin beta, ebulitin gamma, gelonin, gysophilin, hispin, IRAb, IRIP, Iris agglutinin b, Iris ribosome-inactivating protein, JIP60, karasurin-A, KML, lamjapin, luffaculin I, luffin P1, lychnin, MAP, marmorin, ME1, Mirabilis antiviral protein, mistletoe lectin I, mistletoe lectin II, mistletoe lectin III, ML-I, MLI, MLII, MLIII, MOD, modeccin, momorcochin, momorcochin-S, momordin, momordin I, momordin II, momorgrosvin, More, N-glycosidase, n-TCS, neo-trichosanthin, nigrin b, nigritin f1, nigritin f2, PAG, PAGase, PAP, PAP-I, PAP-II, PAP-III, PAP-R, PAP-S, PD-L, PD-L1, PD-L4, pepocin, PGase, pokeweed anti-viral protein, pokeweed antiviral protein, polynucleotide:adenosine glycosidase, polynucleotide:adenosine glycosidase activity, polynucleotide:adenosine glycosylase, porrectin, RA, RCA-II, ribosomal inactivating protein, ribosomal ribonucleate N-glycosidase, ribosome inactivating protein, ribosome-inactivating protein, ribosome-inactivating protein 3, ribosome-inactivating protein type II, ribosome-inactivating proteins, ribosome-inactivating type II protein, ribosome-specific N-glycosidase, ricin, ricin A, ricin A chain, ricin A-chain, ricin toxin A-chain, ricin-A-chain, ricin-like protein, Ricinus communis agglutinin, Ricinus communis agglutinin II, RIP, RIP-1, RIP1, RIP2, RIPII, riproximin, RLP1, RLP2, RLP3, RLP4, RLP5, RLP6, RNA N-glycosidase, RNA-NGA, RNA-specific N-glycosidase, Rpx, rRNA N-glycosidase, RTA, Sambucus nigra agglutinin, Sambucus nigra agglutinin I, SAP, saporin, saporin 6, saporin S-6, saporin S6, saporin-6, saporin-L1, saporin-S6, saporins, SBA, SCO7092, Shiga toxin, Shiga toxin 1, Shiga toxin 1 subunit A, Shiga toxin 2, Shiga toxin 2 subunit A, Shiga toxin type 1, Shiga toxin type 2, Shiga toxin-1, shiga-like toxin, shiga-like toxin I, sieboldin-b, single-chain ribosome-inactivating protein, SLT-I, SLT-II, SLT-IIv, SNA, SNA-I, SNA-V, SnaI, SNAIf, SNAV, SNLRP, SoRIP1, SoRIP2, stenodactylin, Stx, Stx type 1, Stx-1, Stx1, Stx1A, Stx2, Stx2A, Stx2dact, StxB, TCS, trichoanguin, trichomaglin, trichosanthin, Trichosanthrip, trichosnathin, TRIP, tritin-L, tritin-S, TYchi, type 1 ribosome inactivating protein, type 1 ribosome-inactivating protein, type 1 RIP, type 2 ribosome inactivating protein, type 2 ribosome-inactivating protein, type 2 RIP, type 2RIP, type 3 RIP, type I ribosome inactivating protein, type I ribosome-inactivating protein, type I RIP, type II ribosome inactivating protein, type II ribosome-inactivating protein, type II RIP, type III ribosome-inactivating protein, type III RIP, type-1 ribosome-inactivating protein, type-1 RIP, type-2 ribosome-inactivating protein, type-2 RIP, VAA-I, viscinum, Viscum album agglutinin I, viscumin, volkensin

ECTree

     3 Hydrolases
         3.2 Glycosylases
             3.2.2 Hydrolysing N-glycosyl compounds
                3.2.2.22 rRNA N-glycosylase

Purification

Purification on EC 3.2.2.22 - rRNA N-glycosylase

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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 types of ribosome-inactivating proteins: camphorin and cinnamomin
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7 major saporins
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according to Deeks et al., Biochemistry 41, p. 3405-3413, 2002
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affinity chromatography on fetuin-Sepharose 4B and gel filtration of homogenized elderberry bark, homogenized leaves of tobacco plants centrifuged
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affinity column chromatography with a 10 x 2.5 cm column packed with amylose resin specific for maltose-binding protein, eluted pure protein cleaved and dialyzed against 20 mM sodium acetate, pH 5.2, with 0.01% sodium azide, loaded to Sephadex G-100 column
ammonium sulfate precipitation, affinity column chromatography, and Sephadex G-100 gel filtration
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Blue Sepharose 6 column chromatography and CMC-Sephadex C-50 gel filtration
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Blue Sepharose 6 column chromatography, CMC-Sephadex gel filtration, and Sephadex G-50 gel filtration
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Blue Sepharose column chromatography
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by affinity chromatography on a m7G-resin
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Capto Q column chromatography and lactosyl–Sepharose column chromatography
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
centrifugation of homogenized fruiting bodies followed by ion exchange chromatography on a diethylaminoethyl-cellulose column in 10 mM Tris-HCl buffer, pH 7, Affi-gel blue gel column chromatography of unabsorbed protein fraction (with translation-inhibiting activity) in 10 mM Tris-HCl buffer, pH 7 yields bound marmorin
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centrifugation, 0.22 microM filtered, dialyzed against 25 mM sodium chloride in phosphate buffer, pH 6.0, with 2 mM EDTA, application to HP CM FF Sepharose ion exchange column
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CM Sepharose column chromatography and Superdex-75 gel filtration
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fetuin-Sepharose column chromatography and galactose-Sepharose column chromatograhy
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from spring leaf
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Hi-Trap CM-Sepharose column followed by Hi-Trap SP-Sepharose column with 20 mM phosphate buffer, pH 6.7
HisTrap column chromatography
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HisTrap column chromatography and Superdex 200 gel filtration
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homogenized frozen leaves filtered through cheesecloth, centrifuged, saturated with ammonium sulfate to 60%, stirred, all at 4 degrees Celsius, centrifuged, pellet resuspended in 20 mM Tris-HCl, pH 7.5 with 1 mM beta-mercaptoethanol, and 0.2 mM EDTA, dialyzed against buffer, centrifuged, loaded onto HiTrap Q FF column, bound proteins loaded onto HiTrap SP FF column, eluted with NaCl gradient, PAP elutes at about 250 mM NaCl
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immunoaffinity capture with antiricinus communis agglutinin, immobilized and cross-linked to Dynabeads Protein G
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native enzyme 611.4fold from shoots by ammonium sulfate fractionation, ion exchange chromatography and gel filtration
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native enzyme by ammonium sulfate fractionation and gel filtration
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native enzyme from leaves by ion exchange chromatography and gel filtration
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native enzyme from mature seeds by ammonium sulfate fractionation, cation exchange chromatography and gel filtration to 96.6% purity
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native enzyme from seeds
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native enzyme from seeds by ammonium sulfate fractionation, and two steps of gel filtration
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native enzyme from storage roots by immunoaffinity chromatography using a polyclonal anti-ME1 antibody resin
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native gelonin from seeds dialysis of the homogenate, and gel electrophoresis
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native ML-I from plant extract by ammonium sulfate fractionation, and affinity chromatography
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native PAP from spring leaves in four steps, ammonium sulfate fractionation, anion and cation exchange chromatography, and dialysis, to homogeneity
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Ni-NTA column chromatography
particle removal by centrifugation of ground material and concentration by dialysis against PEG
rapid method for extraction and purification from seed
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recombinant B-subunit from Escherichia coli strain BL21(DE3) inclusion bodies to homogeneity
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recombinant functional His6-tagged enzyme, solubilized from Escherichia coli inclusion bodies, by nickel affinity chromatography and gel filtration
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recombinant fusion protein
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recombinant His-tagged RTA and RLPs from Escherichia coli BL21-AI cells by nickel affinity chromatography
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recombinant His-tagged wild-type and mutant PAPs from Escherichia coli strain BL21(DE3) by m7G affinity chromatography
recombinant His6-tagged ME1 from Escherichia coli strain BL21(DE3) by immunoaffinity chromatography, using a polyclonal anti-ME1 antibody resin, and anion exchange chromatography
recombinant His6-tagged optimized abricin A-chain from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant IRAb from transgenic Nicotiana tabacum cv. Samsun plant leaves by ammonium sulfate fractionation, galactose affinity chromatography, hydrophobic interaction chromatography, and dialysis
Iris sp.
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recombinant isozymes from Escherichia coli strain BL21(DE3), purification after overexpression affords refolding by glutathione in refolding buffer, followd by ultrafiltration and cation exchange chromatography
recombinant N-terminally His-tagged isozyme RIP2 from Escherichia coli strain BL21(DE3)
recombinant PAP from Escherichia coli strain JM101 by an alkaline method; recombinant PAP-II from Escherichia coli strain JM101 by an alkaline method; recombinant PAP-R from Escherichia coli strain JM101 by an alkaline method; recombinant PAP-S from Escherichia coli strain JM101 by an alkaline method
recombinant protein
recombinant proteins
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recombinant refolded CBD-intein-Gel fusion protein from inclusion bodies by chitin affinity chromatography with elution using an intein C-terminal self-cleavage method, 96% purity
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recombinant ricin A-chain RTA V81M without N- and C-terminal extensions from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation and anion exchange chromatography
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recombinant secreted ricin-B from tobacco hairy root culture medium
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recombinant SNA-I, SNA-IV, SNA-V, and SNLRP from Nicotiana tabacum cv. Samsun NN transgenic plants by ion exchange chromatography, and fetuin affinity chromatography for SNA-I, recombinant SNA-IV and SNA-V are purified by ammonium sulfate fractionation, galactose affinity chromatography, and hydrophobic interaction chromatography, recombinant SNLRP is purified by ion exchange chromatography and gel filtration
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recombinant Stx1 expressed in Escherichia coli
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recombinant wild-type and mutant ricin A chains from Escherichia coli strain JM101 by anion exchange chromatography and cobalt affinity chromatography to homogeneity
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recombinant wild-type and some mutant RTAs from Escherichia coli strain JM101 by cation exchange chromatography to homogeneity
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sonicated, cleared lysate of cell culture in 20 mM Tris buffer, pH 8.0 with 100 mM NaCl, 1 mM Na-vanadate and protease inhibitors (GST-buffer), incubated with glutathione-Sepharose beads, transferred to column, washed with 0.1 M Tris buffer, pH 7.8 with 0.5 M NaCl, with GST-buffer containing 0.35% Triton X-100, and GST-buffer alone
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wild-type and recombinant proteins
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