1987
DOI: 10.1111/j.1432-1033.1987.tb11210.x
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Purification and immunochemical characterization of the major pertussis‐toxin‐sensitive guanine‐nucleotide‐binding protein of bovine‐neutrophil membranes

Abstract: Bovine peripheral neutrophils contain high levels of a 40-kDa pertussis toxin substrate, which was found highly enriched in a light membrane fraction upon subcellular fractionation of neutrophil homogenates. The 40-kDa pertussis toxin substrate, referred to as a,, was purified to near homogeneity from t h s fraction by sequential ion-exchange, gel-filtration and hydrophobic chromatography. Purified a, was shown to interact with p,, subunits, undergo ADP-ribosylation by pertussis toxin, and bind guanine nucleot… Show more

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Cited by 104 publications
(52 citation statements)
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References 69 publications
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“…Several different G proteins have recently been identified in this cell type (6,15,17) and these are candidates to be tested for their functional role.…”
Section: Discussionmentioning
confidence: 99%
“…Several different G proteins have recently been identified in this cell type (6,15,17) and these are candidates to be tested for their functional role.…”
Section: Discussionmentioning
confidence: 99%
“…1 mM ph,"nylmethylsulfotayl fluoride, and 2/ag/ml soybean trypsin inhibitor) and lysed by nitrogen cavitation as described in [18], except that cells were pressurized with N2 at 4 MPa for 30 mi:, at ~." '7 Removal of unbroken cells and nuclei and preparation of a crude membrane and a cyto~olic fraction from the cavitate was performed as described [18,19].…”
Section: Hemogetff=ation Atzdj'ractionation Of Cellsmentioning
confidence: 99%
“…1 mM ph,"nylmethylsulfotayl fluoride, and 2/ag/ml soybean trypsin inhibitor) and lysed by nitrogen cavitation as described in [18], except that cells were pressurized with N2 at 4 MPa for 30 mi:, at ~." '7 Removal of unbroken cells and nuclei and preparation of a crude membrane and a cyto~olic fraction from the cavitate was performed as described [18,19]. Whole cell ly'~ates were prepared by scraping the cells into ice-cold extraction buffer (20 mM Tris-HCI, pH 7.5:5 mM EDTA, 10 mM EGTA, 37 mM sodium chelate, 3 mM benzamidine, 43 mM 2-mercaptoethanol, and 100 ,tiM phenylmethylsulfon:tl flapride), incubated for 1 h with gentle vortexing every 10 rain, and then centrifuged at 15,000 x g lbr 20 rain.…”
Section: Hemogetff=ation Atzdj'ractionation Of Cellsmentioning
confidence: 99%
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“…An anti-Gi,-common serum (AS/ 6), which was produced against the C-terminal decapeptide (KENLKDCGLF) of rod transducin [17,181, and the antiGIa1 serum (LD/2), which was produced against an internal sequence (LDRIAQPNYI) of Gial [19] were used.…”
Section: Imnzunoprecipitatiori Of G F R O M Plateletsmentioning
confidence: 99%