caa a22 4500 469079495 CHVBK 20180323132945.0 cr unu---uuuuu 170328e19920401xx s 000 0 eng 10.1007/BF00176623 doi (NATIONALLICENCE)springer-10.1007/BF00176623 Lohse Martin Laboratorium für Molekulare Biologie, Universität Müchen, Max-Planck-Institut für Biochemie, W-8033, Martinsried, Germany aut Stable overexpression of human β2-adrenergic receptors in mammalian cells [Elektronische Daten] [Martin Lohse] Summary: Human β2-adrenergic receptors were overexpressed in chinese hamster ovary (CHO) and human epitheloid carcinoma (HeLa) cells. Stable expression in these cells was achieved by transfection of a vector containing the cDNAs for the human β2-adrenergic receptor as well as for dihydrofolate reductase. By stepwise increases of the concentration of methotrexate — an inhibitor of dihydrofolate reductase — the expression in CHO cells could be increased to levels of almost 200 pmol/mg membrane protein, which is more than 1% of the total membrane protein. In contrast, overexpression of the receptors in HeLa cells by the same technique led to cell death. The receptors produced in overexpressing CHO cells were correctly processed and were fully functional with respect to their ligand binding and signalling properties. The adenylyl cyclase activity of membranes from these cells responded with extremely high sensitivity to the β-adrenergic receptor agonist isoproterenol. The receptors could be purified from these membranes to apparent homogeneity by solubilization and chromatography on a single affinity column. Thus, the expression system described here allows the preparation of human β2-adrenergic receptors in quantities sufficient for pharmacological and biochemical investigations. Springer-Verlag, 1992 \2-Adrenergic receptor nationallicence Adenylyl cyclase nationallicence Overexpression nationallicence CHO-cells nationallicence Naunyn-Schmiedeberg's Archives of Pharmacology Springer-Verlag 345/4(1992-04-01), 444-451 0028-1298 345:4<444 1992 345 210 https://doi.org/10.1007/BF00176623 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00176623 text/html Onlinezugriff via DOI NATIONALLICENCE 100 1- Lohse Martin Laboratorium für Molekulare Biologie, Universität Müchen, Max-Planck-Institut für Biochemie, W-8033, Martinsried, Germany aut NATIONALLICENCE 773 0- Naunyn-Schmiedeberg's Archives of Pharmacology Springer-Verlag 345/4(1992-04-01), 444-451 0028-1298 345:4<444 1992 345 210 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer