Br. Conklin et al., SUBSTITUTION OF 3 AMINO-ACIDS SWITCHES RECEPTOR SPECIFICITY OF G(Q)ALPHA TO THAT OF G(I)ALPHA, Nature, 363(6426), 1993, pp. 274-276
AGONIST-BOUND receptors activate heterotrimeric (alphabetagamma) G pro
teins by catalysing replacement of GDP bound to the alpha-subunit by G
TP1-5. Mutations in the C terminus of the alpha-subunit6,7, its covale
nt modification by pertussis toxin-catalysed ribosylation of ADP8, pep
tide-specific antibodies directed against it9-11, and peptides mimicki
ng C-terminal sequences12, all inhibit receptor-mediated activation of
G proteins. The logical prediction-that specific amino-acid residues
at the C-termini of alpha-subunits can determine the abilities of indi
vidual G proteins to discriminate among specific subsets of receptors-
has so far not been tested experimentally. Different hormone receptors
specifically activate G(q) or G(i), whose alpha-subunits (alpha(q) or
alpha(i)) stimulate phosphatidylinositol-specific phospholipase C or
inhibit adenylyl cyclase, respectively1-5. Here we replace C-terminal
amino acids of alpha(q) with the corresponding residues of alpha(i2) t
o create alpha(q)/alpha(i2) chimaeras that can mediate stimulation of
phospholipase C by receptors otherwise coupled exclusively to G(i). A
minimum of three alpha(i2) amino acids, including a glycine three resi
dues from the C terminus, suffices to switch the receptor specificity
of the alpha(q)/alpha(i2) chimaeras. We propose that a C-terminal turn
, centred on this glycine, plays an important part in specifying recep
tor interactions of G proteins in the G(i)/G(o)/G(z) family.