1998
DOI: 10.1021/bi980284o
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Hydrophobicity of Residue351 of the G Protein Gi1α Determines the Extent of Activation by the α2A-Adrenoceptor

Abstract: Cysteine351 is the site for pertussis toxin-catalyzed ADP-ribosylation in the G protein Gi1 alpha. Alteration of this residue, or the equivalent cysteine in other Gi-family G proteins, has been used to examine specific interactions between receptors and these G proteins. However, no systematic analysis has been performed to determine the quantitative effect of such alterations. To address this we mutated cysteine351 of Gi1 alpha to all other possible amino acids. Each of the G protein mutants was transiently c… Show more

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Cited by 86 publications
(151 citation statements)
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References 34 publications
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“…However, mutation of this residue to glycine or isoleucine renders the mutant subunit insensitive to the effects of PTx (34,39). Such mutants have been shown to still functionally interact with receptors as determined by agonist-stimulated 35 S[GTP␥S] binding (37,40,41). We have previously shown that G i␣1 C351G is able to rescue coupling between Kir3.1ϩ3.2A and the transiently transfected A 1 and ␣ 2A receptors in the HKIR3.1/3.2 cell line after PTx treatment (30).…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…However, mutation of this residue to glycine or isoleucine renders the mutant subunit insensitive to the effects of PTx (34,39). Such mutants have been shown to still functionally interact with receptors as determined by agonist-stimulated 35 S[GTP␥S] binding (37,40,41). We have previously shown that G i␣1 C351G is able to rescue coupling between Kir3.1ϩ3.2A and the transiently transfected A 1 and ␣ 2A receptors in the HKIR3.1/3.2 cell line after PTx treatment (30).…”
Section: Resultsmentioning
confidence: 99%
“…Standard molecular cloning and mutagenesis techniques were used throughout. Cell culture, generation of stable cell lines, construction of the bicistronic vector, and point mutations of G ␣ subunits were as described (30,37). For this study we used a similar PCR-based strategy to introduce a C3G mutation at analogous positions in G i␣2 , G i␣3 , and G o␣A .…”
mentioning
confidence: 99%
“…Co-transfection of PTX-insensitive G␣ i1 -C351I, G␣ i2 -C352I, or G␣ i3 -C351I mutants (29) in CCX-CKR-expressing cells abolished the CCL19-induced increase in CRE activity upon PTX pre-treatment (Fig. 7E), which suggests that interaction of G i proteins with CCX-CKR impairs signaling of this receptor to CRE.…”
Section: Ccx-ckr Il2/3 Chimeras Stimulate Campmentioning
confidence: 90%
“…Therefore, to ensure that agonist stimulation of high affinity GTPase activity following expression of the fusion proteins measured guanine nucleotide exchange and hydrolysis by the G protein linked to the GPCR we have used mutants of the G i family G proteins in which the Cys residue which acts as the acceptor for pertussis toxincatalyzed ADP-ribosylation was converted to either Val or Ile (31,34). Pertussis toxin-catalyzed ADP-ribosylation prevents effective interaction between modified G i family G proteins and GPCRs.…”
Section: Discussionmentioning
confidence: 99%
“…In the current study we now demonstrate selective regulation by RGS4 of G i family G proteins coupled to the same GPCR and that the effect of RGS4 includes both concentration dependent increase in rate of agonist-stimulated hydrolysis of GTP and an increase in K m for this nucleotide. (31). The amplicons generated using primers spanning the restriction sites DraI and EcoRI (respectively, in G␣ i1 and in pcDNA3) were subcloned into the ␣ 2A -adrenoreceptor-G␣ i1 fusion construct (20) (20).…”
mentioning
confidence: 99%