PMID: 2117281Aug 1, 1990Paper

Octyl glucoside extracts GTP-binding regulatory proteins from rat brain "synaptoneurosomes" as large, polydisperse structures devoid of beta gamma complexes and sensitive to disaggregation by guanine nucleotides

Proceedings of the National Academy of Sciences of the United States of America
S Nakamura, M Rodbell

Abstract

GTP-binding regulatory proteins are generally purified from cholate-extracted membranes in the form of heterotrimers (G proteins) consisting of a GTP-binding subunit (alpha protein) complexed with a tightly interacted heterodimer termed beta gamma. In this study we extracted the proteins from rat brain "synaptoneurosomes" using the neutral detergent 1-octyl beta-D-glucopyranoside (octyl glucoside). Using specific antibodies for detection by immunoblotting and sucrose gradients for analyzing hydrodynamic properties, we found that each species of alpha protein (alpha subunits of stimulatory, inhibitory, and brain GTP-binding proteins) exhibited a broad range (4 S to greater than 12 S) of polydisperse structures with peak values (5 S to 7 S) considerably greater than that of heterotrimeric G proteins. The beta subunit proteins, for example, appeared as a homogeneous peak at 4.4 S within which only a fraction of the total alpha proteins can be associated. Incubation of octyl glucose extracts at 30 degrees C rapidly sedimented the alpha proteins but not the beta proteins. Incubation at 30 degrees C with guanosine 5'[gamma-thio]triphosphate (10-100 microM) prevented rapid sedimentation. Hydrodynamic analysis revealed that all alpha p...Continue Reading

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Citations

Mar 1, 1994·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·K N Klotz, A J Jesaitis
Jun 1, 1991·Current Opinion in Neurobiology·J Bockaert
Jan 1, 1997·Advances in Enzyme Regulation·M Rodbell
Aug 18, 2000·Immunology and Cell Biology·F S Willard, M F Crouch
Aug 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·S Nakamura, M Rodbell
Nov 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·L JournotY Audigier
Oct 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·S Jahangeer, M Rodbell
Dec 28, 1999·Annual Review of Cell and Developmental Biology·C Montell
Mar 27, 2001·Annual Review of Pharmacology and Toxicology·J Hanoune, N Defer
Apr 1, 1995·Environmental Health Perspectives·M Rodbell
May 5, 1993·Biochemical Pharmacology·G LiapakisK Thermos
Jun 1, 1995·Bioscience Reports·M Rodbell
Jun 1, 1996·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·M OlivottoE Wanke
Nov 2, 2001·The Journal of Biological Chemistry·Valerie A MosserMichael I Schimerlik

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