Reconstitution of cyc− S49 membranes by in vitro translated Gsα
- 17 July 1989
- journal article
- Published by Wiley in FEBS Letters
- Vol. 251 (1-2) , 230-236
- https://doi.org/10.1016/0014-5793(89)81460-7
Abstract
After ADP-ribosylation by cholera toxin which promotes dissociation of the subunits, the α-subunit of Gs(Gsα) remained strongly associated with plasma membranes of wild-type S49 cells, since its interaction with the membrane was insensitive to 1 M KCl. Its association with the membrane was partially disrupted by 6 M urea and totally abolished by treatment with alkali at pH ≥ 11.5. In vitro translated Gsα could interact with plasma membranes from the cyc− mutant of S49 cells as revealed by its cosedimentation with the membrane fraction and incubation of reconstituted membranes with GTPγS did not alter anchorage of Gsα. The characteristics of the association of in vitro translated Gsα with cyc− membranes after GTPγS treatment, i.e. sensitivity to 1 M KCl, 6 M urea and alkali treatment, were very similar to those described for the ADP-ribosylated form in wild-type membranes. Restoration of the coupling between the adrenergic receptor and adenylate cyclase further confirmed the vectorial reconstitution of cyc− membranes by in vitro translated α-subunit of GsKeywords
This publication has 30 references indexed in Scilit:
- Splice Variants of the α Subunit of the G Protein G s Activate Both Adenylyl Cyclase and Calcium ChannelsScience, 1989
- Roles of G protein subunits in transmembrane signallingNature, 1988
- Myristoylated alpha subunits of guanine nucleotide-binding regulatory proteins.Proceedings of the National Academy of Sciences, 1987
- G PROTEINS: TRANSDUCERS OF RECEPTOR-GENERATED SIGNALSAnnual Review of Biochemistry, 1987
- α‐Subunits of Ns are released from the plasma membrane following cholera toxin activationFEBS Letters, 1986
- Molecular Cloning of Complementary DNA for the Alpha Subunit of the G Protein that Stimulates Adenylate CyclaseScience, 1985
- Selection of a Variant Lymphoma Cell Deficient in Adenylate CyclaseScience, 1975
- A highly sensitive adenylate cyclase assayAnalytical Biochemistry, 1974
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Mouse myelomas and lymphomas in cultureExperimental Cell Research, 1970