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ALX-208-002 Revised 22-Jul-08
C3 Exoenzyme
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PRODUCT LINE Signal Transduction
PRODUCT CATEGORY ADP Ribosylation Factor [ARF] / Related Products
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-208-002-C010   10 µg 160.00 USD Add To Cart
ALX-208-002-C050   50 µg 390.00 USD Add To Cart
Product Specification
SOURCE/HOST: Isolated from Clostridium botulinum.
CONCENTRATION: 1mg/ml after reconstitution.
PURITY: ≥98%
APPEARANCE: Lyophilized from 5mM HEPES, pH 8.0.
RECONSTITUTION: Reconstitute with 10µl/50µl distilled water.
SHIPPING: AMBIENT
LONG TERM STORAGE: -20°C
HANDLING: After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles.
Product Description
Exhibits ADP-ribosyltransferase activity. Not neurotoxic. Not cell permeable. The substrate for C3 exoenzyme has been identified as Rho, a GTP-binding Ras-related protein. Rac proteins are approximately 100-fold less active as substrates. The ADP-ribosylation site of exoenzyme C3 on rho A has been identified as Asn41.
Product Specific Literature References
ADP-ribosylation of platelet actin by botulinum C2 toxin: K. Aktories, et al.; Eur. J. Biochem. 161, 155 (1986) Abstract
K. Aktories, et al.; TIPS 8, 158 (1987)
Clostridium botulinum type C produces a novel ADP-ribosyltransferase distinct from botulinum C2 toxin: K. Aktories, et al.; FEBS Lett. 212, 109 (1987) Abstract
Botulinum C2 toxin ADP-ribosylates actin and disorganizes the microfilament network in intact cells: K.H. Reuner, et al.; Eur. J. Cell Biol. 43, 134 (1987) Abstract
Botulinum ADP-ribosyltransferase C3. Purification of the enzyme and characterization of the ADP-ribosylation reaction in platelet membranes: K. Aktories, et al.; Eur. J. Biochem. 172, 445 (1988) Abstract
Functional modification of a 21-kilodalton G protein when ADP- ribosylated by exoenzyme C3 of Clostridium botulinum: E.J. Rubin, et al.; Mol. Cell Biol. 8, 418 (1988) Abstract
The rho gene product expressed in E. coli is a substrate of botulinum ADP-ribosyltransferase C3: K. Aktories, et al.; BBRC 158, 209 (1989) Abstract
K. Aktories and A. Hall; TIPS 10, 415 (1989) Abstract
The mammalian G protein rhoC is ADP-ribosylated by Clostridium botulinum exoenzyme C3 and affects actin microfilaments in Vero cells: P. Chardin, et al.; EMBO J. 8, 1087 (1989) Abstract
Asparagine residue in the rho gene product is the modification site for botulinum ADP-ribosyltransferase: A. Sekine, et al.; J. Biol. Chem. 264, 8602 (1989) Abstract; Full Text
Microinjection of recombinant p21rho induces rapid changes in cell morphology: H.F. Paterson, et al.; J. Cell. Biol. 111, 1001 (1990) Abstract
Characterization of the C3 gene of Clostridium botulinum types C and D and its expression in Escherichia coli: A. Popoff, et al.; Infect. Immun. 59, 3673 (1991) Abstract
The ras protein family: evolutionary tree and role of conserved amino acids: A. Valencia, et al.; Biochemistry 30, 4637 (1991) Abstract
ADP-ribosylation of the ras-related, GTP-binding protein RhoA inhibits lymphocyte-mediated cytotoxicity: P. Lang, et al.; J. Biol. Chem. 267, 11677 (1992) Abstract; Full Text
A rho gene product in human blood platelets. II. Effects of the ADP- ribosylation by botulinum C3 ADP-ribosyltransferase on platelet aggregation: N. Morii, et al.; J. Biol. Chem. 267, 20921 (1992) Abstract; Full Text
The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors: A.J. Ridley, et al.; Cell 70, 389 (1992) Abstract
Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility: K. Takaishi, et al.; Mol. Cell. Biol. 13, 72 (1993) Abstract
Effect of botulinum C3 exoenzyme on cell growth and cytoskeleton organization in transformed human epidermal cells in culture: a possible role for rho protein in epidermal cells: M. Yamamoto, et al.; J. Dermatol. Sci. 8, 103 (1994) Abstract
General Information
All proteins encoded by the superfamily of ras genes are structurally related, bind GTP and exhibit GTPase activity which is regulated by divalent cations. In human cells, three slight variations of the Rho protein (Rho A, B and C, respectively) have been described.
BACKGROUND/TECHNICAL INFORMATION APD-ribosylation is performed as described by Chardin et al. Target G proteins or subcellular fractions corresponding to 50-70µg protein are incubated for 1 hour at 37°C with 20-30ng of C3 in a 10mM HEPES buffer, pH 8.0 containing, 15mM isonicotinic acid hydrazide, 15mM thymidine, 1mM MgCl2, 1mM ATP, 2µM [32P]NAD. The reaction is stopped by addition of 0.06M TRIS-HCl, pH 7.0, 15% (v/v) glycerol, 5% β-mercaptoethanol, 2.3% SDS and 0.001% bromophenol blue. The mixture may then be subjected to SDS-PAGE.
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EnzymesNatural Proteins
 
 

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