Characterization of human 5-HT1 receptors expressed in Sf9 insect cells.
Four human 5-HT receptor subtypes (5-HT1A, 5-HT1D alpha, 5-HT1D beta and 5-HT1E) have been expressed in Sf9 insect cells. All four human 5-hydroxytryptamine receptors produced by Sf9 cells had the expected pharmacological properties. Surprisingly, levels of expression of these receptors were relatively low (1-5 pmol/mg protein). High affinity agonist binding to the four 5-hydroxytryptamine receptors was reduced to different extents by guanine nucleotides and/or NaCl. This suggests that the nature of receptor-G protein coupling and/or the predominant conformational state of the receptors in Sf9 cell membranes varies among the different receptors. Activation of all four receptors inhibited forskolin-stimulated cAMP formation in intact Sf9 cells. Expression of 5-hydroxytryptamine receptors in Sf9 cells should be useful for purification of these receptors, for studies of post-translational modification and for pharmaceutical screening.
Parker
E M
EM
Department of Biophysics, Bristol-Myers Squibb Company, Wallingford, CT 06492.
Grisel
D A
DA
Iben
L G
LG
Nowak
H P
HP
Mahle
C D
CD
Yocca
F D
FD
Gaughan
G T
GT
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D006150:Guanine Nucleotides / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009036:Moths; D011869:Radioligand Assay; D011985:Receptors, Serotonin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017366:Serotonin Receptor Agonists / Q000378:metabolism; D012965:Sodium Chloride / Q000378:metabolism
D006150:Guanine Nucleotides; D011985:Receptors, Serotonin; D011994:Recombinant Proteins; D017366:Serotonin Receptor Agonists; D005576:Colforsin; D012965:Sodium Chloride; D000242:Cyclic AMP; D019204:GTP-Binding Proteins