Identification of the ligand-binding subunit of the human 5-hydroxytryptamine1A
receptor with N-(p-azido-m-125I iodophenethyl)spiperone, a high
affinity radioiodinated photoaffinity probe
J. Raymond, A. Fargin, M. Lohse, J. Regan, S. Senogles, R. Lefkowitz, и M. Caron. Mol Pharmacol, 36 (1):
15-21(июля 1989)Raymond, J R Fargin, A Lohse, M J Regan, J W Senogles, S E Lefkowitz,
R J Caron, M G HL-16037/HL/NHLBI NIH HHS/United States NS-19576/NS/NINDS
NIH HHS/United States S07-RR-05405/RR/NCRR NIH HHS/United States
Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.
United states Molecular pharmacology Mol Pharmacol. 1989 Jul;36(1):15-21..
Аннотация
The ligand-binding subunit of the human 5-hydroxytryptamine1A (5-HT1A)
receptor transiently expressed in COS-7 cells and of the native human
5-HT1A receptor derived from hippocampus and frontal cortex were
identified by photoaffinity labeling with N-(p-azido-m-125Iiodophenethyl)spiperone
( 125IN3-NAPS), previously characterized as a high affinity radioiodinated
D2-dopamine receptor probe. The identity of the ligand-binding subunit
was confirmed by immunoprecipitation with an antipeptide rabbit antiserum,
JWR21, raised against a synthetic peptide derived from the predicted
amino acid sequence of the putative third intracellular loop of the
human 5-HT1A receptor. In transiently transfected COS-7 cells expressing
14 +/- 3 pmol/mg of protein human 5-HT1A receptors, a single broad
75-kDa band was photoaffinity labeled by 125IN3-NAPS. This band
displayed the expected pharmacology of the 5-HT1A receptor, as evidenced
by the ability of a series of competing ligands to block 125IN3-NAPS
photoincorporation. Moreover, antiserum JWR21 specifically and quantitatively
immunoprecipitated the 75-kDa photoaffinity-labeled band from a soluble
extract of the transfected COS-7 cell membranes, further confirming
its identity. Finally, utilizing a combination of photoaffinity labeling
and immunoprecipitation, the native ligand-binding subunit of 62-64
kDa was identified in human hippocampus and frontal cortex. The availability
of the high specific activity, high affinity, photoaffinity ligand
125IN3-NAPS and of a potent immunoprecipitating antiserum (JWR21)
should greatly facilitate the biochemical characterization of the
human 5-HT1A receptor.
Identification of the ligand-binding subunit of the human 5-hydroxytryptamine1A
receptor with N-(p-azido-m-125I iodophenethyl)spiperone, a high
affinity radioiodinated photoaffinity probe
Raymond, J R Fargin, A Lohse, M J Regan, J W Senogles, S E Lefkowitz,
R J Caron, M G HL-16037/HL/NHLBI NIH HHS/United States NS-19576/NS/NINDS
NIH HHS/United States S07-RR-05405/RR/NCRR NIH HHS/United States
Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.
United states Molecular pharmacology Mol Pharmacol. 1989 Jul;36(1):15-21.
%0 Journal Article
%1 Raymond1989
%A Raymond, J. R.
%A Fargin, A.
%A Lohse, M. J.
%A Regan, J. W.
%A Senogles, S. E.
%A Lefkowitz, R. J.
%A Caron, M. G.
%D 1989
%J Mol Pharmacol
%K & 8-Hydroxy-2-(di-n-propylamino)tetralin Affinity Azides/*metabolism Cell D2 Dopamine Dopamine/analysis Humans Iodine Labels/*metabolism Line Precipitin Radioisotopes/diagnostic Serotonin/*analysis/immunology Spiperone/analogs Tests Tetrahydronaphthalenes/metabolism derivatives/*metabolism use Receptor
%N 1
%P 15-21
%T Identification of the ligand-binding subunit of the human 5-hydroxytryptamine1A
receptor with N-(p-azido-m-125I iodophenethyl)spiperone, a high
affinity radioiodinated photoaffinity probe
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2526292
%V 36
%X The ligand-binding subunit of the human 5-hydroxytryptamine1A (5-HT1A)
receptor transiently expressed in COS-7 cells and of the native human
5-HT1A receptor derived from hippocampus and frontal cortex were
identified by photoaffinity labeling with N-(p-azido-m-125Iiodophenethyl)spiperone
( 125IN3-NAPS), previously characterized as a high affinity radioiodinated
D2-dopamine receptor probe. The identity of the ligand-binding subunit
was confirmed by immunoprecipitation with an antipeptide rabbit antiserum,
JWR21, raised against a synthetic peptide derived from the predicted
amino acid sequence of the putative third intracellular loop of the
human 5-HT1A receptor. In transiently transfected COS-7 cells expressing
14 +/- 3 pmol/mg of protein human 5-HT1A receptors, a single broad
75-kDa band was photoaffinity labeled by 125IN3-NAPS. This band
displayed the expected pharmacology of the 5-HT1A receptor, as evidenced
by the ability of a series of competing ligands to block 125IN3-NAPS
photoincorporation. Moreover, antiserum JWR21 specifically and quantitatively
immunoprecipitated the 75-kDa photoaffinity-labeled band from a soluble
extract of the transfected COS-7 cell membranes, further confirming
its identity. Finally, utilizing a combination of photoaffinity labeling
and immunoprecipitation, the native ligand-binding subunit of 62-64
kDa was identified in human hippocampus and frontal cortex. The availability
of the high specific activity, high affinity, photoaffinity ligand
125IN3-NAPS and of a potent immunoprecipitating antiserum (JWR21)
should greatly facilitate the biochemical characterization of the
human 5-HT1A receptor.
@article{Raymond1989,
abstract = {The ligand-binding subunit of the human 5-hydroxytryptamine1A (5-HT1A)
receptor transiently expressed in COS-7 cells and of the native human
5-HT1A receptor derived from hippocampus and frontal cortex were
identified by photoaffinity labeling with N-(p-azido-m-[125I]iodophenethyl)spiperone
[( 125I]N3-NAPS), previously characterized as a high affinity radioiodinated
D2-dopamine receptor probe. The identity of the ligand-binding subunit
was confirmed by immunoprecipitation with an antipeptide rabbit antiserum,
JWR21, raised against a synthetic peptide derived from the predicted
amino acid sequence of the putative third intracellular loop of the
human 5-HT1A receptor. In transiently transfected COS-7 cells expressing
14 +/- 3 pmol/mg of protein human 5-HT1A receptors, a single broad
75-kDa band was photoaffinity labeled by [125I]N3-NAPS. This band
displayed the expected pharmacology of the 5-HT1A receptor, as evidenced
by the ability of a series of competing ligands to block [125I]N3-NAPS
photoincorporation. Moreover, antiserum JWR21 specifically and quantitatively
immunoprecipitated the 75-kDa photoaffinity-labeled band from a soluble
extract of the transfected COS-7 cell membranes, further confirming
its identity. Finally, utilizing a combination of photoaffinity labeling
and immunoprecipitation, the native ligand-binding subunit of 62-64
kDa was identified in human hippocampus and frontal cortex. The availability
of the high specific activity, high affinity, photoaffinity ligand
[125I]N3-NAPS and of a potent immunoprecipitating antiserum (JWR21)
should greatly facilitate the biochemical characterization of the
human 5-HT1A receptor.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Raymond, J. R. and Fargin, A. and Lohse, M. J. and Regan, J. W. and Senogles, S. E. and Lefkowitz, R. J. and Caron, M. G.},
biburl = {https://www.bibsonomy.org/bibtex/2067a2bbf52b10a3c7980784ad97798d9/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {d3f2d5265a89117bb67d17f66b750cbf},
intrahash = {067a2bbf52b10a3c7980784ad97798d9},
issn = {0026-895X (Print) 0026-895X (Linking)},
journal = {Mol Pharmacol},
keywords = {& 8-Hydroxy-2-(di-n-propylamino)tetralin Affinity Azides/*metabolism Cell D2 Dopamine Dopamine/analysis Humans Iodine Labels/*metabolism Line Precipitin Radioisotopes/diagnostic Serotonin/*analysis/immunology Spiperone/analogs Tests Tetrahydronaphthalenes/metabolism derivatives/*metabolism use Receptor},
month = Jul,
note = {Raymond, J R Fargin, A Lohse, M J Regan, J W Senogles, S E Lefkowitz,
R J Caron, M G HL-16037/HL/NHLBI NIH HHS/United States NS-19576/NS/NINDS
NIH HHS/United States S07-RR-05405/RR/NCRR NIH HHS/United States
Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.
United states Molecular pharmacology Mol Pharmacol. 1989 Jul;36(1):15-21.},
number = 1,
pages = {15-21},
shorttitle = {Identification of the ligand-binding subunit of the human 5-hydroxytryptamine1A
receptor with N-(p-azido-m-[125I] iodophenethyl)spiperone, a high
affinity radioiodinated photoaffinity probe},
timestamp = {2010-12-14T18:20:08.000+0100},
title = {Identification of the ligand-binding subunit of the human 5-hydroxytryptamine1A
receptor with N-(p-azido-m-[125I] iodophenethyl)spiperone, a high
affinity radioiodinated photoaffinity probe},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2526292},
volume = 36,
year = 1989
}