Expression of the human GM-CSF receptor α subunit in Saccharomyces cerevisiae Journal Article
| Authors: | Tu, J.; Golde, D. W.; Vera, J. C.; Heaney, M. L. |
| Article Title: | Expression of the human GM-CSF receptor α subunit in Saccharomyces cerevisiae |
| Abstract: | The α subunit of the receptor for human granulocyte-macrophage colony-stimulating factor (GM-CSF) is a 45 kDa membrane protein with a higher apparent molecular weight of 50-85 kDa due to glycosylation. Previously, we had demonstrated that N-glycosylation plays a critical role in the GM-CSF receptor-ligand interaction. To assess the activity of the α subunit of the human GM-CSF receptor (GMRα) in a lower eukaryote, we expressed GMRα in the yeast S. cerevisiae and found that the protein has a lower apparent molecular weight compared with that expressed in mammalian cells. Using indirect immunofluorescence microscopy, we showed that GMRα protein expressed in yeast localizes to the plasma membrane. Although the yeast-expressed GMRα is able to interact with anti GMRα antibody, the heterologously expressed receptor does not bind GM-CSF. Our results indicate that specific sites and/or forms of glycosylation of the GM-CSF receptor are crucial for ligand binding. |
| Keywords: | protein expression; microscopy; nonhuman; protein localization; gene expression; granulocyte macrophage colony stimulating factor; immunofluorescence; saccharomyces cerevisiae; recombinant proteins; cell membrane; glycosylation; yeast; molecular weight; ligand binding; antibody; receptor binding; mammal cell; granulocyte macrophage colony stimulating factor receptor; receptors, granulocyte-macrophage colony-stimulating factor; receptor subunit; gm-csf receptor; n-glycosylation; humans; human; priority journal; article |
| Journal Title: | Cytokines Cellular and Molecular Therapy |
| Volume: | 4 |
| Issue: | 3 |
| ISSN: | 1368-4736 |
| Publisher: | Taylor & Francis Group |
| Date Published: | 1998-09-01 |
| Start Page: | 147 |
| End Page: | 151 |
| Language: | English |
| PUBMED: | 9825839 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 12 December 2016 -- Source: Scopus |
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