Unique isoform of Galpha -interacting protein (RGS-GAIP) selectively discriminates between two Go-mediated pathways that inhibit Ca2+ channels.
Regulators of G-protein signaling (RGS) proteins constitute a large family of GTPase-activating proteins for heterotrimeric G proteins. More than 20 RGS genes have been identified in mammals. One of these, the Galpha-interacting protein (GAIP), preferentially interacts with members of the G(i)/G(o) subfamily of G proteins in mammalian cells, but its selectivity among members of this subfamily in vitro is limited. Here we report the cloning and functional characterization of a unique cDNA isoform of GAIP, derived from embryonic chicken dorsal root ganglion neurons. Chick GAIP is composed of 199 amino acids, organized into a conserved RGS domain (85% identical to human GAIP), and a unique, short N terminus (only 41% identical, 50% homologous to known mammalian orthologues). Consistent with this unique primary structure, chick GAIP has physiological properties that distinguish it from mammalian GAIPs. We have explored the selectivity of chick GAIP in electrophysiological assays of two G(o)-mediated forms of Ca(2+) channel inhibition produced by gamma-aminobutyric acid in chick dorsal root ganglion neurons, voltage-independent inhibition (mediated by G(o)alpha) and voltage-dependent inhibition (mediated by G(o)betagamma). Dialyzing recombinant chick GAIP in these cells selectively reduced voltage-independent inhibition without affecting voltage-dependent inhibition. Mammalian GAIP, tested under identical conditions in previous studies, demonstrated no selectivity between these two inhibitory processes; thus, our results suggest that the functional specificity of chick GAIP is likely to be determined by its unique N terminus.
Tosetti
Patrizia
P
Department of Neuroscience, Tufts University School of Medicine and Molecular Cardiology Research Institute, New England Medical Center, Boston, Massachusetts 02111, USA.
Turner
Timothy
T
Lu
Qiang
Q
Dunlap
Kathleen
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels / Q000502:physiology*; D002642:Chick Embryo; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D020558:GTP Phosphohydrolases / Q000378:metabolism; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D020710:RGS Proteins; D016192:Resting Phase, Cell Cycle; D017386:Sequence Homology, Amino Acid; D005680:gamma-Aminobutyric Acid / Q000502:physiology
D015220:Calcium Channels; D017931:DNA Primers; D018076:DNA, Complementary; D010750:Phosphoproteins; D020033:Protein Isoforms; D020710:RGS Proteins; C097187:regulator of G-protein signalling 19; D005680:gamma-Aminobutyric Acid; D020558:GTP Phosphohydrolases