PMID: 2510152Oct 1, 1989Paper

Isolation of the alpha subunits of GTP-binding regulatory proteins by affinity chromatography with immobilized beta gamma subunits

Proceedings of the National Academy of Sciences of the United States of America
I H Pang, P C Sternweis

Abstract

Immobilized beta gamma subunits of GTP-binding regulatory proteins (G proteins) were used to isolate alpha subunits from solubilized membranes of bovine tissues and to separate specific alpha subunits based on their differential affinities for beta gamma subunits. The beta gamma subunits were cross-linked to omega-aminobutyl agarose. Up to 7 nmol of alpha subunit could bind to each milliliter of beta gamma-agarose and be recovered by elution with AIF4-. This affinity resin effectively separated the alpha subunits of Gi1 and Gi2 from "contaminating" alpha subunits of Go, the most abundant G protein in bovine brain, by taking advantage of the apparent lower affinity of the alpha subunits of Go for beta gamma subunits. The beta gamma-agarose was also used to isolate mixtures of alpha subunits from cholate extracts of membranes from different bovine tissues. alpha subunits of 39-41 kDa (in various ratios) as well as the alpha subunits of Gs were purified. The yields from extracts exceeded 60% for all alpha subunits examined and apparently represented the relative content of alpha subunits in the tissues. This technique can rapidly isolate and identify, from a small amount of sample, the endogenous G proteins in various tissues and ...Continue Reading

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Citations

Jan 1, 1991·Pharmacology & Therapeutics·A Levitzki, A Bar-Sinai
Aug 14, 1992·Biochimica Et Biophysica Acta·S Cockcroft
Oct 29, 1993·Biochimica Et Biophysica Acta·Y Tsunoda
Apr 29, 2005·Brain Research. Molecular Brain Research·Sumita ChakrabartiAlan R Gintzler
Oct 19, 2005·Nature Reviews. Molecular Cell Biology·Craig C Malbon
Aug 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·S M MumbyP C Sternweis
Dec 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·M Strathmann, M I Simon
Sep 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·J A HadwigerR A Firtel
Oct 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·H Kasai
Dec 1, 1992·The Journal of Cell Biology·R SchwaningerW E Balch
Mar 15, 1992·European Journal of Biochemistry·H HeithierE J Helmreich
May 10, 1991·Science·M I SimonN Gautam
Jun 10, 1997·Proceedings of the National Academy of Sciences of the United States of America·C Kleuss, A G Gilman
Apr 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·M E LinderS M Mumby
Sep 18, 2015·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Lutz Birnbaumer
Oct 1, 2015·BioMed Research International·Susana Garcia-Recio, Pedro Gascón
Apr 29, 1992·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·P C SternweisS Gutowski
Oct 26, 1995·Nature·J R MoormanL R Jones
Oct 6, 1997·The Journal of Biological Chemistry·T YamaguchiM Tagaya
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Oct 3, 1999·Physiological Reviews·A J Morris, C C Malbon
Sep 20, 1996·The Journal of Biological Chemistry·J A Duncan, A G Gilman

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