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Literature summary extracted from

  • Craven, R.; Lacy, D.
    Clostridium sordellii lethal-toxin autoprocessing and membrane localization activities drive GTPase glucosylation profiles in endothelial cells (2016), mSphere, 1, e00012-15 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.B62 gene tcsL, recombinant expression of His6-tagged wild-type and mutant enzymes in Bacillus megaterium Paeniclostridium sordellii

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.B62 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
2.4.1.B62 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
2.4.1.B62 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
2.4.1.B62 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

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.
2.4.1.B62 UDP-alpha-D-glucose + a small GTPase Paeniclostridium sordellii
-
 UDP + D-glucosyl-[a small GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + a small GTPase Paeniclostridium sordellii JGS6382
-
 UDP + D-glucosyl-[a small GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac GTPase Paeniclostridium sordellii murine host substrate UDP + D-glucosyl-[Rac GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac GTPase Paeniclostridium sordellii JGS6382 murine host substrate UDP + D-glucosyl-[Rac GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac1 GTPase Paeniclostridium sordellii murine host substrate UDP + D-glucosyl-[Rac1 GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac1 GTPase Paeniclostridium sordellii JGS6382 murine host substrate UDP + D-glucosyl-[Rac1 GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Ral GTPase Paeniclostridium sordellii murine host substrate UDP + D-glucosyl-[Ral GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Ral GTPase Paeniclostridium sordellii JGS6382 murine host substrate UDP + D-glucosyl-[Ral GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rap GTPase Paeniclostridium sordellii murine host substrate UDP + D-glucosyl-[Rap GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rap GTPase Paeniclostridium sordellii JGS6382 murine host substrate UDP + D-glucosyl-[Rap GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Ras GTPase Paeniclostridium sordellii murine host substrate UDP + D-glucosyl-[Ras GTPase]
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.B62 Paeniclostridium sordellii Q46342 Clostridium sordellii
-
2.4.1.B62 Paeniclostridium sordellii JGS6382 Q46342 Clostridium sordellii
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
2.4.1.B62 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)

EC Number Purification (Comment) Organism
2.4.1.B62 recombinant His6-tagged wild-type and mutant enzymes from Bacillus megaterium by nickel affinity chromatography and gel filtration Paeniclostridium sordellii

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.B62 UDP-alpha-D-glucose + a small GTPase
-
 Paeniclostridium sordellii UDP + D-glucosyl-[a small GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + a small GTPase
-
 Paeniclostridium sordellii JGS6382 UDP + D-glucosyl-[a small GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac GTPase murine host substrate Paeniclostridium sordellii UDP + D-glucosyl-[Rac GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac GTPase murine host substrate Paeniclostridium sordellii JGS6382 UDP + D-glucosyl-[Rac GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac1 GTPase murine host substrate Paeniclostridium sordellii UDP + D-glucosyl-[Rac1 GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rac1 GTPase murine host substrate Paeniclostridium sordellii JGS6382 UDP + D-glucosyl-[Rac1 GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Ral GTPase murine host substrate Paeniclostridium sordellii UDP + D-glucosyl-[Ral GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Ral GTPase murine host substrate Paeniclostridium sordellii JGS6382 UDP + D-glucosyl-[Ral GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rap GTPase murine host substrate Paeniclostridium sordellii UDP + D-glucosyl-[Rap GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Rap GTPase murine host substrate Paeniclostridium sordellii JGS6382 UDP + D-glucosyl-[Rap GTPase]
-
 ?
2.4.1.B62 UDP-alpha-D-glucose + Ras GTPase murine host substrate Paeniclostridium sordellii UDP + D-glucosyl-[Ras GTPase]
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.B62 lethal-toxin
-
 Paeniclostridium sordellii
2.4.1.B62 TcsL
-
 Paeniclostridium sordellii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.B62 33 37 assay at Paeniclostridium sordellii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.B62 7.4
-
 assay at Paeniclostridium sordellii

General Information

EC Number General Information Comment Organism
2.4.1.B62 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
2.4.1.B62 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