Expression in animal cells of the 5-HT1A receptor by a vaccinia virus vector system.
The co-infection or infection-transfection variants of the T7 RNA polymerase/vaccinia vector system were used to express 5-HT1ARs in COS-7, BSC-40 and GH3 cells, with co-infection giving ca. 3-fold higher level than infection-transfection. Binding affinities were similar to those of the endogenous 5-HT1AR, with highest affinities for 5-HT and 8-OH-DPAT. Functional properties were demonstrated by assays of agonist-stimulated GTPase activity and its inhibition by pertussin toxin. Immunoblot assays showed expression of the unglycosylated and glycosylated receptor protein in the membrane and, surprisingly, in the cytosolic fractions.
Ho
B Y
BY
Division of Biology, California Institute of Technology, Pasadena 91125.
Karschin
A
A
Raymond
J R
JR
Branchek
T
T
Lester
H A
HA
Davidson
N
N
D017371:8-Hydroxy-2-(di-n-propylamino)tetralin; D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D015870:Gene Expression; D005814:Genes, Viral; D005822:Genetic Vectors; D011985:Receptors, Serotonin / Q000235:genetics* / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism; D013604:T-Phages / Q000201:enzymology; D013764:Tetrahydronaphthalenes / Q000378:metabolism; D014616:Vaccinia virus / Q000235:genetics*
D011985:Receptors, Serotonin; D013764:Tetrahydronaphthalenes; D012701:Serotonin; D017371:8-Hydroxy-2-(di-n-propylamino)tetralin; D012321:DNA-Directed RNA Polymerases; D020558:GTP Phosphohydrolases