PMID: 3106040May 15, 1987

Purification and immunochemical characterization of the major pertussis-toxin-sensitive guanine-nucleotide-binding protein of bovine-neutrophil membranes

European Journal of Biochemistry
P GierschikK H Jakobs

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 alpha n, was purified to near homogeneity from this fraction by sequential ion-exchange, gel-filtration and hydrophobic chromatography. Purified alpha n was shown to interact with beta gamma subunits, undergo ADP-ribosylation by pertussis toxin, and bind guanine nucleotides with high affinity. The mobility of purified alpha n on SDS/polyacrylamide gels was intermediate between those of the alpha subunits of Gi and Go, purified from bovine brain, and slightly lower than the mobility of the alpha subunit of transducin (Gt). Several polyclonal antisera against the alpha subunits of bovine Gt and Go did not react with alpha n on immunoblots. CW 6, a polyclonal antiserum reactive against the bovine alpha i, reacted only minimally with alpha n. These results suggest that the major pertussis toxin substrate of bovine neutrophils, designated Gn, is structurally different from previously identified pertussis toxin substrates and may represent a novel guanine-nucleotide-binding pro...Continue Reading

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Citations

Jul 1, 1994·Journal of Cellular Biochemistry·L P WennogleS I Hu
Feb 1, 1993·Journal of Neuroscience Research·G GimplH Kettenmann
Jan 1, 1989·Immunologic Research·R G Coffey
Oct 1, 1988·Immunology Today·M M Harnett, G G Klaus
May 1, 1989·Molecular and Cellular Endocrinology·U RudolphW Rosenthal
Mar 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·M R PhilipsG Weissmann
Dec 1, 1988·The Journal of Cell Biology·O Nüsse, M Lindau
Nov 20, 1989·European Journal of Biochemistry·E HerrmannK H Jakobs
Jun 15, 1992·European Journal of Biochemistry·M CampsP Gierschik
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Related Concepts

Adenosine Diphosphate Ribose
Bos indicus
Cell Fractionation
Plasma Membrane
Guanosine Triphosphate
Peroxidase-Antiperoxidase Complex Technique
Neutrophil Band Cells
Agglutinogen 2, Bordetella Pertussis
Thionucleotides
Guanosine 5'-O-(3-Thiotriphosphate)

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