A highly conserved glycine within linker I and the extreme C terminus of G protein alpha subunits interact cooperatively in switching G protein-coupled receptor-to-effector specificity
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A highly conserved glycine within linker I and the extreme C terminus of G protein alpha subunits interact cooperatively in switching G protein-coupled receptor-to-effector specificity. / Kostenis, Evi; Martini, Lene; Ellis, James; Waldhoer, Maria; Heydorn, Arne; Rosenkilde, Mette M; Norregaard, Pia K; Jorgensen, Rasmus; Whistler, Jennifer L; Milligan, Graeme.
I: Journal of Pharmacology and Experimental Therapeutics, Bind 313, Nr. 1, 2004, s. 78-87.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - A highly conserved glycine within linker I and the extreme C terminus of G protein alpha subunits interact cooperatively in switching G protein-coupled receptor-to-effector specificity
AU - Kostenis, Evi
AU - Martini, Lene
AU - Ellis, James
AU - Waldhoer, Maria
AU - Heydorn, Arne
AU - Rosenkilde, Mette M
AU - Norregaard, Pia K
AU - Jorgensen, Rasmus
AU - Whistler, Jennifer L
AU - Milligan, Graeme
N1 - Keywords: Amino Acid Sequence; Animals; Blotting, Western; COS Cells; Calcium; Cell Membrane; Cercopithecus aethiops; Conserved Sequence; DNA; Enzyme-Linked Immunosorbent Assay; GTP-Binding Protein alpha Subunits; Glycine; Guanosine 5'-O-(3-Thiotriphosphate); Inositol Phosphates; Ligands; Molecular Sequence Data; Receptors, G-Protein-Coupled; Signal Transduction; Transfection
PY - 2004
Y1 - 2004
N2 - Numerous studies have attested to the importance of the extreme C terminus of G protein alpha subunits in determining their selectivity of receptor recognition. We have previously reported that a highly conserved glycine residue within linker I is important for constraining the fidelity of receptor recognition by Galpha(q) proteins. Herein, we explored whether both modules (linker I and extreme C terminus) interact cooperatively in switching G protein-coupled receptor (GPCR)-to-effector specificity and created as models mutant Galpha(q) proteins in which glycine was replaced with various amino acids and the C-terminal five Galpha(q) residues with the corresponding Galpha(i) or Galpha(s) sequence. Coupling properties of the mutated Galpha(q) proteins were determined after coexpression with a panel of 13 G(i)-and G(s) -selective receptors and compared with those of Galpha proteins modified in only one module. Galpha proteins modified in both modules are significantly more efficacious in channeling non-G(q) -selective receptors to G(q)-mediated signaling events compare with those containing each module alone. Additive effects of both modules were observed even if individual modules lacked an effect on GPCR-to-effector specificity. Dually modified Galpha proteins were also superior in conferring high-affinity agonist sites onto a coexpressed GPCR in the absence, but not in the presence, of guanine nucleotides. Together, our data suggest that receptor-G protein coupling selectivity involves cooperative interactions between the extreme C terminus and linker I of Galpha proteins and that distinct determinants of selectivity exist for individual receptors.
AB - Numerous studies have attested to the importance of the extreme C terminus of G protein alpha subunits in determining their selectivity of receptor recognition. We have previously reported that a highly conserved glycine residue within linker I is important for constraining the fidelity of receptor recognition by Galpha(q) proteins. Herein, we explored whether both modules (linker I and extreme C terminus) interact cooperatively in switching G protein-coupled receptor (GPCR)-to-effector specificity and created as models mutant Galpha(q) proteins in which glycine was replaced with various amino acids and the C-terminal five Galpha(q) residues with the corresponding Galpha(i) or Galpha(s) sequence. Coupling properties of the mutated Galpha(q) proteins were determined after coexpression with a panel of 13 G(i)-and G(s) -selective receptors and compared with those of Galpha proteins modified in only one module. Galpha proteins modified in both modules are significantly more efficacious in channeling non-G(q) -selective receptors to G(q)-mediated signaling events compare with those containing each module alone. Additive effects of both modules were observed even if individual modules lacked an effect on GPCR-to-effector specificity. Dually modified Galpha proteins were also superior in conferring high-affinity agonist sites onto a coexpressed GPCR in the absence, but not in the presence, of guanine nucleotides. Together, our data suggest that receptor-G protein coupling selectivity involves cooperative interactions between the extreme C terminus and linker I of Galpha proteins and that distinct determinants of selectivity exist for individual receptors.
U2 - 10.1124/jpet.104.080424
DO - 10.1124/jpet.104.080424
M3 - Journal article
C2 - 15615862
VL - 313
SP - 78
EP - 87
JO - Journal of Pharmacology and Experimental Therapeutics
JF - Journal of Pharmacology and Experimental Therapeutics
SN - 0022-3565
IS - 1
ER -
ID: 15530576