Characterization of adenosine A1 receptor in a cell line (28A) derived
from rabbit collecting tubule
W. Spielman, K. Klotz, L. Arend, B. Olson, D. LeVier, и U. Schwabe. Am J Physiol, 263 (2 Pt 1):
C502-8(августа 1992)Spielman, W S Klotz, K N Arend, L J Olson, B A LeVier, D G Schwabe,
U R01 DK-39654/DK/NIDDK NIH HHS/United States Research Support, U.S.
Gov't, P.H.S. United states The American journal of physiology Am
J Physiol. 1992 Aug;263(2 Pt 1):C502-8..
Аннотация
We have previously reported that in several renal cell types, adenosine
receptor agonists inhibit adenylyl cyclase and activate phospholipase
C via a pertussis toxin-sensitive G protein. In the present study,
in 28A cells, both of these adenosine receptor-mediated responses
were inhibited by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), a highly
selective A1 adenosine receptor antagonist. The binding characteristics
of the adenosine A1 receptor in the 28A renal cell line were studied
using the radiolabeled antagonist 3HDPCPX to determine whether
two separate binding sites could account for these responses. Saturation
binding of 3HDPCPX to 28A cell membranes revealed a single class
of A1 binding sites with an apparent Kd value of 1.4 nM and maximal
binding capacity of 64 fmol/mg protein. Competition experiments with
a variety of adenosine agonists gave biphasic displacement curves
with a pharmacological profile characteristic of A1 receptors. Comparison
of 3HDPCPX competition binding data from 28A cell membranes with
rabbit brain membranes, a tissue with well-characterized A1 receptors,
reveals that the A1 receptor population in 28A cells has similar
agonist binding affinities to the receptor population in brain but
has a considerably lower density. Addition of guanosine 5'-triphosphate
(100 microM) to 28A cell membranes caused the competition curves
to shift from biphasic to monophasic, indicating that the A1 receptors
exist in two interconvertible affinity states because of their coupling
to G proteins.(ABSTRACT TRUNCATED AT 250 WORDS)
Spielman, W S Klotz, K N Arend, L J Olson, B A LeVier, D G Schwabe,
U R01 DK-39654/DK/NIDDK NIH HHS/United States Research Support, U.S.
Gov't, P.H.S. United states The American journal of physiology Am
J Physiol. 1992 Aug;263(2 Pt 1):C502-8.
%0 Journal Article
%1 Spielman1992
%A Spielman, W. S.
%A Klotz, K. N.
%A Arend, L. J.
%A Olson, B. A.
%A LeVier, D. G.
%A Schwabe, U.
%D 1992
%J Am J Physiol
%K & AMP/antagonists Adenosine/analogs Adenylate Animals Binding, Bordetella/pharmacology Brain/metabolism Calcium/metabolism Cell Collecting/cytology/*metabolism Competitive Cyclase Cyclic Factors, Kidney Line Medulla/metabolism Membrane/metabolism Pertussis Purinergic/classification/drug Rabbits Toxin Tubules, Virulence Xanthines/pharmacology derivatives/pharmacology effects/*metabolism inhibitors Receptor
%N 2 Pt 1
%P C502-8
%T Characterization of adenosine A1 receptor in a cell line (28A) derived
from rabbit collecting tubule
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1325120
%V 263
%X We have previously reported that in several renal cell types, adenosine
receptor agonists inhibit adenylyl cyclase and activate phospholipase
C via a pertussis toxin-sensitive G protein. In the present study,
in 28A cells, both of these adenosine receptor-mediated responses
were inhibited by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), a highly
selective A1 adenosine receptor antagonist. The binding characteristics
of the adenosine A1 receptor in the 28A renal cell line were studied
using the radiolabeled antagonist 3HDPCPX to determine whether
two separate binding sites could account for these responses. Saturation
binding of 3HDPCPX to 28A cell membranes revealed a single class
of A1 binding sites with an apparent Kd value of 1.4 nM and maximal
binding capacity of 64 fmol/mg protein. Competition experiments with
a variety of adenosine agonists gave biphasic displacement curves
with a pharmacological profile characteristic of A1 receptors. Comparison
of 3HDPCPX competition binding data from 28A cell membranes with
rabbit brain membranes, a tissue with well-characterized A1 receptors,
reveals that the A1 receptor population in 28A cells has similar
agonist binding affinities to the receptor population in brain but
has a considerably lower density. Addition of guanosine 5'-triphosphate
(100 microM) to 28A cell membranes caused the competition curves
to shift from biphasic to monophasic, indicating that the A1 receptors
exist in two interconvertible affinity states because of their coupling
to G proteins.(ABSTRACT TRUNCATED AT 250 WORDS)
@article{Spielman1992,
abstract = {We have previously reported that in several renal cell types, adenosine
receptor agonists inhibit adenylyl cyclase and activate phospholipase
C via a pertussis toxin-sensitive G protein. In the present study,
in 28A cells, both of these adenosine receptor-mediated responses
were inhibited by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), a highly
selective A1 adenosine receptor antagonist. The binding characteristics
of the adenosine A1 receptor in the 28A renal cell line were studied
using the radiolabeled antagonist [3H]DPCPX to determine whether
two separate binding sites could account for these responses. Saturation
binding of [3H]DPCPX to 28A cell membranes revealed a single class
of A1 binding sites with an apparent Kd value of 1.4 nM and maximal
binding capacity of 64 fmol/mg protein. Competition experiments with
a variety of adenosine agonists gave biphasic displacement curves
with a pharmacological profile characteristic of A1 receptors. Comparison
of [3H]DPCPX competition binding data from 28A cell membranes with
rabbit brain membranes, a tissue with well-characterized A1 receptors,
reveals that the A1 receptor population in 28A cells has similar
agonist binding affinities to the receptor population in brain but
has a considerably lower density. Addition of guanosine 5'-triphosphate
(100 microM) to 28A cell membranes caused the competition curves
to shift from biphasic to monophasic, indicating that the A1 receptors
exist in two interconvertible affinity states because of their coupling
to G proteins.(ABSTRACT TRUNCATED AT 250 WORDS)},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Spielman, W. S. and Klotz, K. N. and Arend, L. J. and Olson, B. A. and LeVier, D. G. and Schwabe, U.},
biburl = {https://www.bibsonomy.org/bibtex/2e5e054ecf11736ae051f543128f65a97/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {317a05054b12a170847de9f3539e5a31},
intrahash = {e5e054ecf11736ae051f543128f65a97},
issn = {0002-9513 (Print) 0002-9513 (Linking)},
journal = {Am J Physiol},
keywords = {& AMP/antagonists Adenosine/analogs Adenylate Animals Binding, Bordetella/pharmacology Brain/metabolism Calcium/metabolism Cell Collecting/cytology/*metabolism Competitive Cyclase Cyclic Factors, Kidney Line Medulla/metabolism Membrane/metabolism Pertussis Purinergic/classification/drug Rabbits Toxin Tubules, Virulence Xanthines/pharmacology derivatives/pharmacology effects/*metabolism inhibitors Receptor},
month = Aug,
note = {Spielman, W S Klotz, K N Arend, L J Olson, B A LeVier, D G Schwabe,
U R01 DK-39654/DK/NIDDK NIH HHS/United States Research Support, U.S.
Gov't, P.H.S. United states The American journal of physiology Am
J Physiol. 1992 Aug;263(2 Pt 1):C502-8.},
number = {2 Pt 1},
pages = {C502-8},
shorttitle = {Characterization of adenosine A1 receptor in a cell line (28A) derived
from rabbit collecting tubule},
timestamp = {2010-12-14T18:20:09.000+0100},
title = {Characterization of adenosine A1 receptor in a cell line (28A) derived
from rabbit collecting tubule},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=1325120},
volume = 263,
year = 1992
}