Prosaposin receptor: evidence for a G-protein-associated receptor
| ISSN: | 0006-291X |
|---|---|
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::95108afb8f94253ac001d33217c72ab9 https://pubmed.ncbi.nlm.nih.gov/9388493 |
| Rights: | CLOSED |
| Přírůstkové číslo: | edsair.doi.dedup.....95108afb8f94253ac001d33217c72ab9 |
| Autor: | Masao Hiraiwa, John S. O'Brien, W M Campana, Brian M. Martin |
| Rok vydání: | 1997 |
| Předmět: |
GTP'
G protein Biophysics Receptors Cell Surface Biology Pertussis toxin Biochemistry Chromatography Affinity Saposins Receptors G-Protein-Coupled Mice Neuroblastoma Affinity chromatography GTP-Binding Proteins Tumor Cells Cultured Animals Amino Acid Sequence Nerve Growth Factors Receptor Molecular Biology Peptide sequence G alpha subunit Glycoproteins Prosaposin Cell Membrane Brain Cell Biology Molecular biology Peptide Fragments Molecular Weight Guanosine 5'-O-(3-Thiotriphosphate) Electrophoresis Polyacrylamide Gel Papio |
| Zdroj: | Biochemical and biophysical research communications. 240(2) |
| ISSN: | 0006-291X |
| Popis: | Prosaposin, the precursor of sphingolipid activator protein (saposins A-D), has been identified as a neurotrophic factor capable of inducing neural differentiation and preventing cell death. The putative prosaposin receptor was partially purified from baboon brain membranes by affinity chromatography using a saposin C-column. The purified preparation gave a single major protein band with an apparent molecular weight of 54 kDa on SDS-PAGE. Affinity cross-linking of 11 kDa 125I-saposin C demonstrated the presence of a 66 kDa product, indicative of an apparent molecular weight of 55 kDa for the receptor. A GTP gamma S-binding assay using cell membranes from SHSY5Y neural cells demonstrated agonist stimulated binding of [35S]-GTP gamma S upon treatment with prosaptide TX14(A) a peptide from the neurotrophic region; maximal binding was obtained at 2 nM. TX14(A) stimulated binding was abolished by prior treatment of SHSY5Y cells with pertussis toxin and by a scrambled and an all D-amino acid-derivative of the 14-mer. A 14-mer mutant prosaptide (6N-->6D) competed with TX14(A) with a Ki of 0.7 nM. Immunoblot analysis using an antibody against the G0 alpha subunit demonstrated that the purified receptor preparation contained a 40 kDa reactive band consistent with association of G0 alpha and the receptor. These findings indicate that the signaling induced by prosaposin and TX14(A) is generated by binding to a G0-protein associated receptor. |
| Databáze: | OpenAIRE |
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