Any feedback?
Literature summary for 3.2.2.22 extracted from
- The effect of mutations surrounding and within the active site on the catalytic activity of ricin A chain (2004), Eur. J. Biochem., 271, 153-162.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of wild-type and mutant RTAs in Escherichia coli strain JM101 | Ricinus communis |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant mutant RTAs N122A and R213D, sitting drop vapour diffusion method using microbridges, X-ray diffraction structure determination and analysis at 1.4 A and 1.9 A resolution, respectively | Ricinus communis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D75A | site-directed mutagenesis of the RTA residue, the mutant RTA shows very low expression levels so that purification to homogeneity is not achieved | Ricinus communis |
| D75N | site-directed mutagenesis of the RTA residue, the mutant RTA shows very low expression levels so that purification to homogeneity is not achieved | Ricinus communis |
| D75S | site-directed mutagenesis of the RTA residue, the mutant RTA shows very low expression levels so that purification to homogeneity is not achieved | Ricinus communis |
| N122A | site-directed mutagenesis of the RTA residue, 37.5fold reduced activity compared to the wild-type RTA, the reconstituted enzyme comprising RTB and N122A RTA shows about 30fold reduced cytotoxicity compared to the wild-type enzyme | Ricinus communis |
| N78S | site-directed mutagenesis of the RTA residue, less than 2fold reduced activity compared to the wild-type RTA, the reconstituted enzyme comprising RTB and N78S RTA shows about 2fold reduced cytotoxicity compared to the wild-type enzyme | Ricinus communis |
| R134A | site-directed mutagenesis of the RTA residue, the expression of the recombinant mutant is abolished, R134 probably plays a structural role | Ricinus communis |
| R134Q | site-directed mutagenesis of the RTA residue, the expression of the recombinant mutant is abolished, R134 probably plays a structural role | Ricinus communis |
| R213A | site-directed mutagenesis of the RTA residue, the mutant RTA shows unaltered structure, but 10fold reduced activity compared to the wild-type RTA | Ricinus communis |
| R213D | site-directed mutagenesis of the RTA residue, the mutant RTA shows unaltered structure, but 2fold reduced activity compared to the wild-type RTA | Ricinus communis |
| R258A | site-directed mutagenesis of the RTA residue, similar activity compared to the wild-type RTA | Ricinus communis |
| R258D | site-directed mutagenesis of the RTA residue, slightly reduced activity compared to the wild-type RTA | Ricinus communis |
| R48A | site-directed mutagenesis of the RTA residue, slightly reduced activity compared to the wild-type RTA | Ricinus communis |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | - |
Ricinus communis |  |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Ricinus communis | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and some mutant RTAs from Escherichia coli strain JM101 by cation exchange chromatography to homogeneity | Ricinus communis |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| A4324 in 28S rRNA + H2O = deadenylated 28S rRNA + adenine | active site structure and catalytic mechanism, residue R213 is important for catalysis | Ricinus communis |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| reassociation of purified recombinant RTAs mixed with RTB in presence of 0.1 M lactose and 2% v/v 2-mercaptoethanol by dialysis for 24 h against 1 l of NaCl/phosphate solution containing 0.1 M lactose, followed by further 36 h against 5 l of NaCl/phosphate solution | Ricinus communis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 26S rRNA + H2O | depurination of ribosomes from yeast, product determination using aniline coupling | Ricinus communis | ? | - |
? | |
| additional information | binding of RTA to the GAGA tetraloop of rRNA, interaction analysis | Ricinus communis | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | heterodimer formed by a catalytic subunit A, RTA and a carbohydrate binding subunit B, RTB | Ricinus communis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ricin | - |
Ricinus communis |
| type II ribosome-inactivating protein | - |
Ricinus communis |
| type II RIP | - |
Ricinus communis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
N-glycosidase assay at | Ricinus communis |
| 37 | - |
in vivo cytotoxicity assay at | Ricinus communis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.6 | - |
assay at | Ricinus communis |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
