Heterotrimeric and small molecular mass GTP-binding proteins of rat brain neuronal and glial nuclei.

Abstract

Heterotrimeric and small molecular mass guanine nucleotide binding (GTP-binding) proteins were found in neuronal and glial nuclei isolated from rat brain. Neuronal nuclei bound 0.213 +/- 0.055 pmoles of GTP/microg protein (n = 8) and glial nuclei bound 0.145 +/- 0.038 pmoles of GTP/microg protein (n = 8). The intrinsic GTPase activity of neuronal and glial nuclei was 0.0019 +/- 0.0005 pmoles GTP hydrolyzed/min/microg protein (n = 10) and 0.0022 +/- 0.0006 pmoles GTP hydrolyzed/min/microg protein (n = 10), respectively. Western blot analysis was carried out using a peptide-specific antibody that recognizes a common sequence in the alpha-subunit of the various heterotrimeric G-proteins. The antibody revealed the presence of a polypeptide of molecular mass of 40 kDa only in neuronal nuclei. Small molecular mass GTP-binding proteins were detected by incubating nitrocellulose blots with [alpha-32P]GTP. The results demonstrated the presence of 25-26 kDa GTP-binding proteins in both populations of nuclei. However, the binding of [alpha-32P]GTP to neuronal nuclei was approximately 3-fold greater than to the glial nuclei. Further analysis by two-dimensional polyacrylamide gel electrophoresis resolved the neuronal nuclei 26 kDa protein into three forms (a-c) with the most acidic form (c) being the major species. The neuronal 25 kDa protein was resolved into two forms that were present in approximately equal concentration. In glial nuclei, only the 26 kDa (c) and a small amount of the 25 kDa proteins were detected. However, both populations of nuclei contained the small molecular mass GTP-binding protein, ran. Differential association of non-ran small molecular mass GTP-binding proteins and heterotrimeric G-proteins with neuronal nuclei suggests a potential role for these guanine nucleotide binding proteins in the function of this cell type.