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Literature summary for 2.4.1.B62 extracted from
- Clostridium sordellii lethal-toxin autoprocessing and membrane localization activities drive GTPase glucosylation profiles in endothelial cells (2016), mSphere, 1, e00012-15 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene tcsL, recombinant expression of His6-tagged wild-type and mutant enzymes in Bacillus megaterium | Paeniclostridium sordellii |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C698A | site-directed mutagenesis of the autoprocessing domain, mutant TcsL C698A is able to quickly glucosylate Rac1, similar to wild-type TcsL, but is attenuated in its ability to glucosylate Ras GTPases. The introduction of the autoprocessing mutation does not impact the glucosylation of Rac1 or H-Ras in an in vitro assay | Paeniclostridium sordellii |
| F17N/R18A | site-directed mutagenesis in the GTD membrane localization domain, on the surface of the membrane localization domain (MLD), the mutant shows a defect in membrane association in a liposome binding assay | Paeniclostridium sordellii |
| F17N/R18A/C698A | site-directed mutagenesis in the GTD membrane localization domain, on the surface of the membrane localization domain (MLD), the mutant shows a defect in membrane association in a liposome binding assay. The triple mutant is also inhibited in both Rac1 and Ras glucosylation | Paeniclostridium sordellii |
| additional information | when glucosyltransferase-deficient TcsL mutant, TcsL DxD, is used to intoxicate cells, the loss of the glucosyltransferase activity renders the mutant unable to glucosylate both Rac1 and Ras | Paeniclostridium sordellii |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| UDP-alpha-D-glucose + a small GTPase | Paeniclostridium sordellii | - |
UDP + D-glucosyl-[a small GTPase] | - |
? |  |
| UDP-alpha-D-glucose + a small GTPase | Paeniclostridium sordellii JGS6382 | - |
UDP + D-glucosyl-[a small GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac GTPase | Paeniclostridium sordellii | murine host substrate | UDP + D-glucosyl-[Rac GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac GTPase | Paeniclostridium sordellii JGS6382 | murine host substrate | UDP + D-glucosyl-[Rac GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac1 GTPase | Paeniclostridium sordellii | murine host substrate | UDP + D-glucosyl-[Rac1 GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac1 GTPase | Paeniclostridium sordellii JGS6382 | murine host substrate | UDP + D-glucosyl-[Rac1 GTPase] | - |
? | |
| UDP-alpha-D-glucose + Ral GTPase | Paeniclostridium sordellii | murine host substrate | UDP + D-glucosyl-[Ral GTPase] | - |
? | |
| UDP-alpha-D-glucose + Ral GTPase | Paeniclostridium sordellii JGS6382 | murine host substrate | UDP + D-glucosyl-[Ral GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rap GTPase | Paeniclostridium sordellii | murine host substrate | UDP + D-glucosyl-[Rap GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rap GTPase | Paeniclostridium sordellii JGS6382 | murine host substrate | UDP + D-glucosyl-[Rap GTPase] | - |
? | |
| UDP-alpha-D-glucose + Ras GTPase | Paeniclostridium sordellii | murine host substrate | UDP + D-glucosyl-[Ras GTPase] | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Paeniclostridium sordellii | Q46342 | Clostridium sordellii | - |
| Paeniclostridium sordellii JGS6382 | Q46342 | Clostridium sordellii | - |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| proteolytic modification | maturation of the host cell endosome causes a conformational change in the pore-forming domain of TcsL, causing it to form a pore in the endosomal membrane. The autoprocessing domain is activated by host inositol hexakisphosphate and cleaves the glucosyltransferase domain (GTD), presumably to permit access to substrates residing at the plasma membrane | Paeniclostridium sordellii |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His6-tagged wild-type and mutant enzymes from Bacillus megaterium by nickel affinity chromatography and gel filtration | Paeniclostridium sordellii |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| UDP-alpha-D-glucose + a small GTPase | - |
Paeniclostridium sordellii | UDP + D-glucosyl-[a small GTPase] | - |
? | |
| UDP-alpha-D-glucose + a small GTPase | - |
Paeniclostridium sordellii JGS6382 | UDP + D-glucosyl-[a small GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac GTPase | murine host substrate | Paeniclostridium sordellii | UDP + D-glucosyl-[Rac GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac GTPase | murine host substrate | Paeniclostridium sordellii JGS6382 | UDP + D-glucosyl-[Rac GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac1 GTPase | murine host substrate | Paeniclostridium sordellii | UDP + D-glucosyl-[Rac1 GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rac1 GTPase | murine host substrate | Paeniclostridium sordellii JGS6382 | UDP + D-glucosyl-[Rac1 GTPase] | - |
? | |
| UDP-alpha-D-glucose + Ral GTPase | murine host substrate | Paeniclostridium sordellii | UDP + D-glucosyl-[Ral GTPase] | - |
? | |
| UDP-alpha-D-glucose + Ral GTPase | murine host substrate | Paeniclostridium sordellii JGS6382 | UDP + D-glucosyl-[Ral GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rap GTPase | murine host substrate | Paeniclostridium sordellii | UDP + D-glucosyl-[Rap GTPase] | - |
? | |
| UDP-alpha-D-glucose + Rap GTPase | murine host substrate | Paeniclostridium sordellii JGS6382 | UDP + D-glucosyl-[Rap GTPase] | - |
? | |
| UDP-alpha-D-glucose + Ras GTPase | murine host substrate | Paeniclostridium sordellii | UDP + D-glucosyl-[Ras GTPase] | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| lethal-toxin | - |
Paeniclostridium sordellii |
| TcsL | - |
Paeniclostridium sordellii |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 33 | 37 | assay at | Paeniclostridium sordellii |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.4 | - |
assay at | Paeniclostridium sordellii |
General Information
| General Information | Comment | Organism |
|---|---|---|
| malfunction | the difference in cellular Rac1 and Ras glucosylation when the autoprocessing activity is ablated (mutation C698A) suggests that there is a localization difference for Rac1 and Ras. Mutations in the GTD membrane localization domain inhibit TcsL cytotoxicity | Paeniclostridium sordellii |
| physiological function | maturation of the host cell endosome causes a conformational change in the pore-forming domain of TcsL, causing it to form a pore in the endosomal membrane. The autoprocessing domain is activated by host inositol hexakisphosphate and cleaves the glucosyltransferase domain (GTD), presumably to permit access to substrates residing at the plasma membrane. The GTD glucosylates small GTPases, predominately Rac, Ras, Ral, and Rap. The glucosylation leads to cytoskeletal rearrangement and rounding of the cells and also causes the induction of apoptosis. Lethal-toxin TcsL shows cytotoxicity in murine pulmonary microvascular endothelial cells (mPMVECs) as model cells, incubation at 33°C or 37°C. Cell viability is determined by CellTiter-Glo luciferase 24 h after intoxication. TcsL autoprocessing and glucosyltransferase activities are important for cytotoxicity, TcsL cytotoxicity is dependent upon GTD localization to the cell membrane. The membrane localization domain (MLD) interaction with the membrane is important for the glucosylation of both Rac1 and Ras | Paeniclostridium sordellii |
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