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AB318256

Recombinant Clostridioides difficile Toxin B/TcdB Protein (His-tag)

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Recombinant Clostridioides difficile Toxin B/TcdB Protein (His-tag) is a Clostridium difficile Fragment protein, in the 2 to 543 aa range, expressed in Escherichia coli, with >95%, <0.005 EU/µg endotoxin level, suitable for Mass Spec.

View Alternative Names

toxB, tcdB, Toxin B

2 Images
Mass Spectrometry - Recombinant Clostridioides difficile Toxin B/TcdB Protein (His-tag) (AB318256)
  • Mass Spec

Supplier Data

Mass Spectrometry - Recombinant Clostridioides difficile Toxin B/TcdB Protein (His-tag) (AB318256)

Mass determination by ESI-TOF. Predicted MW is 64640.00 Da (+/- 10 Da by ESI-TOF). Observed MW is 64510.51 Da.

SDS-PAGE - Recombinant Clostridioides difficile Toxin B/TcdB Protein (His-tag) (AB318256)
  • SDS-PAGE

Supplier Data

SDS-PAGE - Recombinant Clostridioides difficile Toxin B/TcdB Protein (His-tag) (AB318256)

SDS-PAGE analysis of ab318256 under reducing conditions for 2ug protein.

Key facts

Purity

>95% HPLC

Endotoxin level

<0.005 EU/µg

Expression system

Escherichia coli

Tags

His tag C-Terminus

Applications

Mass Spec

applications

Biologically active

No

Mass Spectrometry

LC-MS/MS

Accession

P18177

Animal free

Yes

Carrier free

Yes

Species

Clostridium difficile

Storage buffer

pH: 8 Constituents: 1.4586% Sodium chloride, 0.7505% Tris HCl

storage-buffer

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Complementary Products

Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

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Primary Antibodies

AB16502

Anti-NF-kB p65 antibody

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NF-kB p65 antibody (AB16502)

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Primary Antibodies

AB209845

Anti-GSDMD antibody [EPR19828]

RabMAb|Recombinant|KO Validated

Immunoprecipitation - Anti-GSDMD antibody [EPR19828] (AB209845)

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Primary Antibodies

AB133462

Anti-IKB alpha (phospho S36) antibody [EPR6235(2)]

20ul selling size|RabMAb|Recombinant

Western blot - Anti-IKB alpha (phospho S36) antibody [EPR6235(2)] (AB133462)

  • 4
  • 1
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Mass Spec": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Sequence info

[{"sequence":"SLVNRKQLEKMANVRFRTQEDEYVAILDALEEYHNMSENTVVEKYLKLKDINSLTDIYIDTYKKSGRNKALKKFKEYLVTEVLELKNNNLTPVEKNLHFVWIGGQINDTAINYINQWKDVNSDYNVNVFYDSNAFLINTLKKTVVESAINDTLESFRENLNDPRFDYNKFFRKRMEIIYDKQKNFINYYKAQREENPELIIDDIVKTYLSNEYSKEIDELNTYIEESLNKITQNSGNDVRNFEEFKNGESFNLYEQELVERWNLAAASDILRISALKEIGGMYLDVDMLPGIQPDLFESIEKPSSVTVDFWEMTKLEAIMKYKEYIPEYTSEHFDMLDEEVQSSFESVLASKSDKSEIFSSLGDMEASPLEVKIAFNSKGIINQGLISVKDSYCSNLIVKQIENRYKILNNSLNPAISEDNDFNTTTNTFIDSIMAEANADNGRFMMELGKYLRVGFFPDVKTTINLSGPEAYAAAYQDLLMFKEGSMNIHLIEADLRNFEISKTNISQSTEQEMASLWSFDDARAKAQFEEYKRNYFEGSL","proteinLength":"Fragment","predictedMolecularWeight":"64.64 kDa","actualMolecularWeight":"64.51 kDa","aminoAcidEnd":543,"aminoAcidStart":2,"nature":"Recombinant","expressionSystem":null,"accessionNumber":"P18177","tags":[{"tag":"His","terminus":"C-Terminus"}]}]
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Properties and storage information

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-80°C
Appropriate long-term storage conditions
-80°C
False
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Clostridium difficile Toxin B also known as CD Toxin B or B protein is a potent toxin produced by the bacterium Clostridium difficile. It is one of the major virulence factors alongside Toxin A. This protein carries significant weight approximately 270 kDa. It is expressed in the intestinal tract when Clostridium difficile colonizes and proliferates often leading to severe gastrointestinal conditions. The presence of toxins including both Toxin A and Toxin B contributes to the pathogenicity of C. diff infection.
Biological function summary

This toxin exerts its effects by modifying intracellular signaling pathways disrupting tight junctions and leading to cell apoptosis. Toxin B specifically targets the Rho family of GTPases through glucosylation leading to actin cytoskeleton disorganization and subsequent cell rounding and tissue damage. It functions independently but works in conjunction with Toxin A to enhance virulence. As part of its biological role Toxin B proves essential in the pathogenesis of diseases associated with C. diff infection.

Pathways

Research shows Toxin B's involvement in the disruption of the actin cytoskeleton pathway. This pathway alteration results from direct modification of small GTP-binding proteins such as RhoA Rac and Cdc42. Toxin B's activity leads to loss of cell structure and increased cell lysis showcasing how it fits into major cellular integrity pathways. Its interaction with these proteins places it alongside other microbial toxins that manipulate host cell signaling.

Toxin B has a strong association with Clostridium difficile infection (CDI) which often manifests as pseudomembranous colitis. CDI presents with symptoms such as severe diarrhea and colonic inflammation. Furthermore Toxin B contributes significantly to the disease severity in comparison to other microbial toxins. While Toxin A also plays a role studies suggest that Toxin B's impact on the intestinal epithelium is more severe marking it as a critical target for therapeutic intervention against CDI.
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Specifications

Form

Liquid

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General info

Function

Toxin B. Precursor of a cytotoxin that targets and disrupts the colonic epithelium, inducing the host inflammatory and innate immune responses and resulting in diarrhea and pseudomembranous colitis (PubMed : 20844489, PubMed : 24919149). TcdB constitutes the main toxin that mediates the pathology of C.difficile infection, an opportunistic pathogen that colonizes the colon when the normal gut microbiome is disrupted (PubMed : 19252482, PubMed : 20844489). Compared to TcdA, TcdB is more virulent and more important for inducing the host inflammatory and innate immune responses (PubMed : 19252482, PubMed : 24919149). This form constitutes the precursor of the toxin : it enters into host cells and mediates autoprocessing to release the active toxin (Glucosyltransferase TcdB) into the host cytosol (PubMed : 10768933, PubMed : 11152463, PubMed : 12941936, PubMed : 17334356, PubMed : 20498856). Targets colonic epithelia by binding to the frizzled receptors FZD1, FZD2 and FZD7, and enters host cells via clathrin-mediated endocytosis (PubMed : 27680706). Frizzled receptors constitute the major host receptors in the colonic epithelium, but other receptors, such as CSPG4 or NECTIN3/PVRL3, have been identified (PubMed : 25547119, PubMed : 26038560, PubMed : 27680706). Binding to carbohydrates and sulfated glycosaminoglycans on host cell surface also contribute to entry into cells (By similarity). Once entered into host cells, acidification in the endosome promotes the membrane insertion of the translocation region and formation of a pore, leading to translocation of the GT44 and peptidase C80 domains across the endosomal membrane (PubMed : 11152463, PubMed : 12941936, PubMed : 24567384). This activates the peptidase C80 domain and autocatalytic processing, releasing the N-terminal part (Glucosyltransferase TcdB), which constitutes the active part of the toxin, in the cytosol (PubMed : 17334356, PubMed : 27571750).. Glucosyltransferase TcdB. Active form of the toxin, which is released into the host cytosol following autoprocessing and inactivates small GTPases (PubMed : 16157585, PubMed : 17901056, PubMed : 24905543, PubMed : 24919149, PubMed : 7777059, PubMed : 8144660). Acts by mediating monoglucosylation of small GTPases of the Rho family (Rac1, RhoA, RhoB, RhoC, RhoG and Cdc42) in host cells at the conserved threonine residue located in the switch I region ('Thr-37/35'), using UDP-alpha-D-glucose as the sugar donor (PubMed : 16157585, PubMed : 17901056, PubMed : 24905543, PubMed : 24919149, PubMed : 7777059). Monoglucosylation of host small GTPases completely prevents the recognition of the downstream effector, blocking the GTPases in their inactive form, leading to actin cytoskeleton disruption and cell death, resulting in the loss of colonic epithelial barrier function (PubMed : 24919149, PubMed : 7777059).

Sequence similarities

Belongs to the clostridial glucosylating toxin (LCGT) family.

Post-translational modifications

Toxin B. Undergoes autocatalytic cleavage to release the N-terminal part (Glucosyltransferase TcdB), which constitutes the active part of the toxin, in the host cytosol (PubMed:12941936, PubMed:17334356, PubMed:27571750). 1D-myo-inositol hexakisphosphate-binding (InsP6) activates the peptidase C80 domain and promotes autoprocessing (PubMed:17334356).

Subcellular localisation

Host endosome membrane

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Product protocols

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Target data

Toxin B. Precursor of a cytotoxin that targets and disrupts the colonic epithelium, inducing the host inflammatory and innate immune responses and resulting in diarrhea and pseudomembranous colitis (PubMed : 20844489, PubMed : 24919149). TcdB constitutes the main toxin that mediates the pathology of C.difficile infection, an opportunistic pathogen that colonizes the colon when the normal gut microbiome is disrupted (PubMed : 19252482, PubMed : 20844489). Compared to TcdA, TcdB is more virulent and more important for inducing the host inflammatory and innate immune responses (PubMed : 19252482, PubMed : 24919149). This form constitutes the precursor of the toxin : it enters into host cells and mediates autoprocessing to release the active toxin (Glucosyltransferase TcdB) into the host cytosol (PubMed : 10768933, PubMed : 11152463, PubMed : 12941936, PubMed : 17334356, PubMed : 20498856). Targets colonic epithelia by binding to the frizzled receptors FZD1, FZD2 and FZD7, and enters host cells via clathrin-mediated endocytosis (PubMed : 27680706). Frizzled receptors constitute the major host receptors in the colonic epithelium, but other receptors, such as CSPG4 or NECTIN3/PVRL3, have been identified (PubMed : 25547119, PubMed : 26038560, PubMed : 27680706). Binding to carbohydrates and sulfated glycosaminoglycans on host cell surface also contribute to entry into cells (By similarity). Once entered into host cells, acidification in the endosome promotes the membrane insertion of the translocation region and formation of a pore, leading to translocation of the GT44 and peptidase C80 domains across the endosomal membrane (PubMed : 11152463, PubMed : 12941936, PubMed : 24567384). This activates the peptidase C80 domain and autocatalytic processing, releasing the N-terminal part (Glucosyltransferase TcdB), which constitutes the active part of the toxin, in the cytosol (PubMed : 17334356, PubMed : 27571750).. Glucosyltransferase TcdB. Active form of the toxin, which is released into the host cytosol following autoprocessing and inactivates small GTPases (PubMed : 16157585, PubMed : 17901056, PubMed : 24905543, PubMed : 24919149, PubMed : 7777059, PubMed : 8144660). Acts by mediating monoglucosylation of small GTPases of the Rho family (Rac1, RhoA, RhoB, RhoC, RhoG and Cdc42) in host cells at the conserved threonine residue located in the switch I region ('Thr-37/35'), using UDP-alpha-D-glucose as the sugar donor (PubMed : 16157585, PubMed : 17901056, PubMed : 24905543, PubMed : 24919149, PubMed : 7777059). Monoglucosylation of host small GTPases completely prevents the recognition of the downstream effector, blocking the GTPases in their inactive form, leading to actin cytoskeleton disruption and cell death, resulting in the loss of colonic epithelial barrier function (PubMed : 24919149, PubMed : 7777059).
See full target information tcdB
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