Rapid purification of Bordetella pertussis toxin by alternating affinity and hydrophobic chromatography

Svoboda, Michal; Hannecart-Pokorni, E.; Borremans, Martine; Christophe, Jean

doi:10.1016/0003-2697(86)90360-x

Cited by 21 publications

(7 citation statements)

References 26 publications

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“…Cultures were diluted 1:40 in 20 ml of syncase broth and grown for 2.5 h at 37 °C on a shaker at 120 rpm; mitomycin C (Sigma, St. Louis, MO, USA) was next added to a final concentration of 0.5 μg/ml, and cultivated for 16 h. Cells were concentrated by centrifugation; the supernatant was passed through 0.22-μm pore filters, and analysed by chromatography. ArtABs (except for ArtAB-Sb) were purified by consecutive elution from Affi-Gel Blue (Bio-Rad, Hercules, CA, USA) and hydroxyapatite columns followed by hydrophobic interaction chromatography (HIC), as previously described for Ptx purification 38 . Desalting or buffer exchange of protein solutions was performed using a PD-10 Column (GE Healthcare).…”

Section: Methodsmentioning

confidence: 99%

Characterization of pertussis-like toxin from Salmonella spp. that catalyzes ADP-ribosylation of G proteins

Tamamura

Tanaka

Uchida

2017

Sci Rep

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Salmonella Typhimurium definitive phage type (DT) 104 produces a pertussis-like toxin (ArtAB-DT104), which catalyzes ADP-ribosylation of pertussis toxin sensitive G proteins. However, the prevalence of ArtAB and its toxicity have not been established. We report here that, in addition to DT104, S. Worthington, and S. bongori, produce ArtAB homologs, designated ArtAB-SW and ArtAB-Sb, respectively. We purified and characterized these ArtAB toxins, which comprise a 27-kDa A subunit (ArtA) and 13.8-kDa pentameric B subunits (ArtB). While the sequence of the A subunit, which is ADP-ribosyltransferase, is similar to the A subunit sequences of other ArtABs, the B subunit of ArtAB-Sb is divergent compared to the B subunit sequences of other ArtABs. Intraperitoneal injection of purified ArtABs was fatal in mice; the 50% lethal doses of ArtAB-DT104 and ArtAB-SW were lower than that of ArtAB-Sb, suggesting that ArtB plays an influential role in the toxicity of ArtABs. ArtABs catalyzed ADP-ribosylation of G proteins in RAW 264.7 murine macrophage-like cells, and increased intracellular cyclic AMP levels. ArtAB-DT104 and ArtAB-SW, but not ArtAB-Sb, stimulated insulin secretion in mice; however, unlike Ptx, ArtABs did not induce leukocytosis. This disparity in biological activity may be explained by differences in ADP-ribosylation of target G proteins.

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Section: Methodsmentioning

confidence: 99%

Characterization of pertussis-like toxin from Salmonella spp. that catalyzes ADP-ribosylation of G proteins

Tamamura

Tanaka

Uchida

2017

Sci Rep

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“…This inhibitory effect of CCK and gastrin was observed in membranes from a transformed rat pancreatic acinar cell line but not in membranes from normal rat pancreas where, in contrast, the enzymatic activity was stimulated by CCK. This absence of CCK inhibition in normal pancreas could not be attributed to a lack of Gi since this organ contains a 41 kDa protein that can be ADPribosylated in the presence of Bordetella pertussis toxin [4]. Our results are, therefore, best explained by considering distinct populations of CCK receptors in AR 4-2J cells and in normal pancreas.…”

Section: Discussionmentioning

confidence: 57%

“…Pertussis toxin, prepared as described [4], was kindly given by Dr M. Svoboda (Dept of Biochemistry and Nutrition, Medical School, Universit6 Libre de Bruxelles) and, when used, was incorporated into the culture medium at a concentration of 0.1/~g/ml, 18 h before membrane preparation.…”

Section: Methodsmentioning

confidence: 99%

CCK and gastrin inhibit adenylate cyclase activity through a pertussis toxin‐sensitive mechanism in the tumoral rat pancreatic acinar cell line AR 4‐2J

et al. 1988

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(Thr28,Nleat)CCK(23-33) (CCK-9) and gastrin(1-17)l (gastrin) inhibited adenylate cyclase activity in membranes from the tumoral rat pancreatic acinar cell line AR 4-2J through a Bordetella pertussis toxin-sensitive mechanism. This contrasted with the stimulatory effect exerted by CCK-9 on adenylate cyclase activity in membranes from normal rat pancreas. The relative potency of CCK-9, gastrin, and related peptides in inhibiting adenylate cyclase, when confronted with previous evidence, suggests that 'non-selective CCK-gastrin CCK-B receptors' predominating over 'selective CCK-A receptors' in the AR 4-2J cell line, favored the coupling of the first receptors to adenylate cyclase through G, while CCK-A receptors capable of stimulating the enzyme through Gs were detected only after Bordetella pertussis toxin pretreatment.

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“…CCK can also stimulate PC hydrolysis by phospholipase D, contributing to DAG formation [2]; to our knowledge, the role of a Gprotein in the coupling of CCK receptors to phospholipase D has not yet been investigated. [3] are detected by ADP-ribosylation in the presence of CT. Low-affinity CCK receptors, whose occupancy correlates with the downstroke of the dose-response curve for enzyme secretion, are also coupled through G, with adenylate cyclase in the rat pancreas [4,5]. Pancreatic adenylate cyclase is submitted to a negative regulation through somatostatin receptors [6] coupled to adenylate cyclase through Gi [7] already identified by ADP-ribosylation in the presence of PT [8].…”

Section: Introductionmentioning

confidence: 99%

Novel GTP‐binding proteins in plasma membranes and zymogen granule membranes from rat pancreas and in pancreatic AR 4‐2J cell membranes

1990

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Rapid purification of Bordetella pertussis toxin by alternating affinity and hydrophobic chromatography

Cited by 21 publications

References 26 publications

Characterization of pertussis-like toxin from Salmonella spp. that catalyzes ADP-ribosylation of G proteins

Characterization of pertussis-like toxin from Salmonella spp. that catalyzes ADP-ribosylation of G proteins

CCK and gastrin inhibit adenylate cyclase activity through a pertussis toxin‐sensitive mechanism in the tumoral rat pancreatic acinar cell line AR 4‐2J

Novel GTP‐binding proteins in plasma membranes and zymogen granule membranes from rat pancreas and in pancreatic AR 4‐2J cell membranes

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