Heterogeneity of three electrophoretically distinct G_o α‐subunits in mammalian brain

Abstract

So far three splice variants of the a:o-gen¢ coding for two qo proteins have been identified by molecular cloning, and the corresponding proteins purified, In the preen( study subtype-specific peptide antibodies revealed the existence of an electrophoretically distinct third form of ~o in mammalian brain membranes which migrates more slowly on SDS-PAGE and shows a more acidic pl value than the other ~to-subtmits. Each of the three ao-subunits is detected as two isol'orms when enriched from brain membranes, Rodent ao-subunits differ from non-rodent species in their electrophoretic mobilities, The results suggest that (it there may exist a novel go-subunit which reacts with an aot-subunit-slX'Cific antibody, (ii) each %-subunit may exist in more than one co-or posttran~lationally modified iso form in brain membranes, and (iii)differences betw~n ao-subunit..~ from different species exist which are detectable by gel electrophorctic methods.

Phenotypic spectrum of GNAO1 variants: epileptic encephalopathy to involuntary movements with severe developmental delay

“…Gαo1 is the most abundant Gαo protein in the mammalian brain. 14,15 Of the two patients, both showed severe intellectual disability and motor developmental delay, indicating that the affected Gαo1 function could cause neurological impairments. Although one of the two patients (patient 3 in this study) showed no seizures, another showed intractable seizures from early neonatal period.…”

Receptors couple to L‐type calcium channels via distinct Go proteins in rat neuroendocrine cell lines

“…Purities of Gá isoforms, GiÏo mixtures and Gâã complexes were greater than 95% as determined by silver staining of 10% SDS-polyacrylamide gels. The purified PTXsensitive Gá isoforms were identified by immunoreactivity to subtype-specific polyclonal antibodies Spicher, N urnberg, J ager, Rosenthal & Schultz, 1992;N urnberg et al 1994) and proved to be devoid of contamination by other PTX-sensitive Gá subunits, as assessed by autoradiographic analysis after PTXmediated 32 P-ADP ribosylation; standard protocols were applied for PTX-mediated ADP ribosylation (N urnberg, 1997).…”

“…Separating gels contained 6 Ò urea and 9% (wÏv) acrylamide. Immunoblotting was performed as described, and filter-bound antibodies were visualized by a colour reaction catalysed by goat-anti-rabbit-IgG coupled to peroxidase (dilution, 1:1000; Sigma) and the enhanced chemiluminescence (ECL) Western-blotting detection system (Amersham) (Spicher et al 1992). ECL-stained blots were exposed to X-ray films for 1-20 min.…”

“…ECL-stained blots were exposed to X-ray films for 1-20 min. Antiserum AS 6 (anti-áo,common) was raised in rabbits against the peptide (c)nlkedgisaakdvk which corresponds to the predicted amino acids 22-35 of either Gáo sequence (Spicher et al 1992). Specificities of antisera AS 6, AS 201 and AS 248 for distinct recombinant á-subunits expressed in E. coli were tested as detailed previously Spicher et al , 1992.…”

Purification of the G-protein G13 from rat brain membranes

“…In order to avoid smeared protein bands, samples obtained from flyagarose affinity chromatography were subjected to sample preparation as described by Schagger and von Jagow [18]. Immunoblotting was performed as described previously [19]. Filter …”