Abstract
Antibodies generated to a synthetic decapeptide, RMHLRQYELL, representing the carboxyl-terminus of Gs-α have been characterized in immunoblots and functional studies. This antibody, designated RM, reacts exclusively with a doublet of proteins of 52 and 45 kDa in immunoblots of bovine brain and wild-type S49 murine lymphoma cell membranes. No such reactivity is seen in membranes from cyc- S49 cells, which lack Gs. RM blocks receptor-mediated activation of Gs and adenylyl cyclase in membranes from wild-type S49 cells. RM could also immunoprecipitate adenylyl cyclase activity in detergent extracts from GTP[γ]S- or fluoride-preactivated bovine brain membranes; thus binding of αs to effector and carboxyl-terminal antibody was mutually compatible. Such experiments provide an approach for the elucidation of functionally relevant interactions of G-proteins with receptors and electors in the membrane.
Original language | English (US) |
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Pages (from-to) | 189-194 |
Number of pages | 6 |
Journal | FEBS Letters |
Volume | 249 |
Issue number | 2 |
DOIs | |
State | Published - Jun 5 1989 |
Externally published | Yes |
Keywords
- Adenylyl cyclase
- Adrenergic receptor, β-
- GTP-binding protein
- Immunoprecipitation
ASJC Scopus subject areas
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology