ADP-ribosylation and de-ADP-ribosylation of the rho protein by Clostridium botulinum exoenzyme C3. Regulation by EDTA, guanine nucleotides and pH

Habermann, Barbara; Mohr, Christiane; Just, Ingo; Aktories, Klaus

doi:10.1016/0167-4838(91)90537-a

Cited by 17 publications

(5 citation statements)

References 31 publications

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“…The binding of nicotinamide was too weak to be detected despite an experimental pH that was close to the optima for both the forward and reverse enzyme reactions (Habermann et al, 1991). However, soaking with 5 mM NAD or ADP-ribose caused crystal disintegration.…”

Section: Ligand Binding and Crystal Packingmentioning

confidence: 90%

C3 exoenzyme fromClostridium botulinum: structure of a tetragonal crystal form and a reassessment of NAD-induced flexure

Evans

Holloway

Sutton³

et al. 2004

Acta Crystallogr D Biol Cryst

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C3 exoenzyme from Clostridium botulinum (C3bot1) ADP-ribosylates and thereby inactivates Rho A, B and C GTPases in mammalian cells. The structure of a tetragonal crystal form has been determined by molecular replacement and re®ned to 1.89 A Ê resolution. It is very similar to the apo structures determined previously from two different monoclinic crystal forms. An objective reassessment of available apo and nucleotide-bound C3bot1 structures indicates that, contrary to a previous report, the protein possesses a rigid core formed largely of-strands and that the general¯exure that accompanies NAD binding is concentrated in two peripheral lobes. Tetragonal crystals disintegrate in the presence of NAD, most likely because of disruption of essential crystal contacts.

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Section: Ligand Binding and Crystal Packingmentioning

confidence: 90%

C3 exoenzyme fromClostridium botulinum: structure of a tetragonal crystal form and a reassessment of NAD-induced flexure

Evans

Holloway

Sutton³

et al. 2004

Acta Crystallogr D Biol Cryst

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“…To test the influence of PAO on the structure of RhoA, RhoA was subjected to C 3 lim -catalyzed [ 32 P]ADP-ribosylation, because C 3 lim exclusively modifies native RhoA (Habermann et al, 1991). RhoA from cells, treated with 50 M PAO (Fig.…”

Section: Downloaded Frommentioning

confidence: 99%

Thiol-Modifying Phenylarsine Oxide Inhibits Guanine Nucleotide Binding of Rho but Not of Rac GTPases

et al. 2003

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Phenylarsine oxide (PAO) is a phosphotyrosine phosphatase inhibitor that cross-links vicinal thiol groups, thereby inactivating phosphatases possessing XCysXXCysX motifs. The RhoAGTPase, but not the Rac1-GTPase, also possesses vicinal cysteines within the guanine nucleotide-binding region (aa [13][14][15][16][17][18][19][20] and the phosphohydrolase activity site. Treatment of Caco-2 cells with PAO showed a dose-dependent reorganization of the actin cytoskeleton, indicating involvement of Rho GTPases. As tested by pull-down experiments, RhoA, but not Rac1, from cell lysates was inactivated by PAO in a concentration-dependent manner. Modification of RhoA by PAO resulted in altered mobility on SDS-polyacrylamide gel electrophoresis, and PAO-modified RhoA was no longer substrate for C3-catalyzed ADP-ribosylation. Furthermore, RhoA treated with PAO, but not Rac1 treated with PAO, lost its property to bind to guanine nucleotides. Matrix-assisted laser desorption ionization-mass analysis of PAO-modified RhoA showed a mass shift according to an adduction of a single PAO molecule per molecule RhoA. Further analysis of Glu-C-generated RhoA peptides confirmed binding of PAO to a peptide harboring the guanine nucleotide binding region. Thus, PAO does not exclusively inhibit phosphotyrosine phosphatases but also inactivates RhoA by alteration of nucleotide binding.

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“…All C3 exoenzymes recognize RhoA, B, or C; and S. aureus EDIN also modifies RhoE [24]. Rho-GDP is a preferred substrate for C3 [32], as Rho-Asn41 is solvent-accessible in the GDP-bound structure [33]. In contrast, the Asn41 residue of Rho found in a Rho-GDI complex is hidden and thus resistant to C3-mediated ADP-ribosylation [34].…”

Section: Structure and Enzymatic Activitymentioning

confidence: 91%

ADP-ribosylating toxins modifying the actin cytoskeleton

Barth

Stiles

Popoff

2015

The Comprehensive Sourcebook of Bacterial Protein Toxins

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ADP-ribosylation and de-ADP-ribosylation of the rho protein by Clostridium botulinum exoenzyme C3. Regulation by EDTA, guanine nucleotides and pH

Cited by 17 publications

References 31 publications

C3 exoenzyme fromClostridium botulinum: structure of a tetragonal crystal form and a reassessment of NAD-induced flexure

C3 exoenzyme fromClostridium botulinum: structure of a tetragonal crystal form and a reassessment of NAD-induced flexure

Thiol-Modifying Phenylarsine Oxide Inhibits Guanine Nucleotide Binding of Rho but Not of Rac GTPases

ADP-ribosylating toxins modifying the actin cytoskeleton

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