Involvement of Asn-293 in stereospecific agonist recognition and
in activation of the beta 2-adrenergic receptor
K. Wieland, H. Zuurmond, C. Krasel, A. Ijzerman, and M. Lohse. Proc Natl Acad Sci U S A, 93 (17):
9276-81(August 1996)Wieland, K Zuurmond, H M Krasel, C Ijzerman, A P Lohse, M J Comparative
Study Research Support, Non-U.S. Gov't United states Proceedings
of the National Academy of Sciences of the United States of America
Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9276-81..
Abstract
To investigate the molecular mechanism for stereospecific binding
of agonists to beta 2-adrenergic receptors we used receptor models
to identify potential binding sites for the beta-OH-group of the
ligand, which defines the chiral center. Ser-165, located in transmembrane
helix IV, and Asn-293, situated in the upper half of transmembrane
helix VI, were identified as potential binding sites. Mutation of
Ser-165 to Ala did not change the binding of either isoproterenol
isomer as revealed after transient expression in human embryonic
kidney (HEK)-293 cells. In contrast, a receptor mutant in which Asn-293
was replaced by Leu showed substantial loss of stereospecific isoproterenol
binding. Adenylyl cyclase stimulation by this mutant after stable
expression in CHO cells confirmed the substantial loss of stereospecificity
for isoproterenol. In a series of agonists the loss of affinity in
the Leu-293 mutant receptor was strongly correlated with the intrinsic
activity of the compounds. Full agonists showed a 10-30-fold affinity
loss, whereas partial agonists had almost the same affinity for both
receptors. Stereospecific recognition of antagonists was unaltered
in the Leu-293 mutant receptor. These data indicate a relationship
between stereospecificity and intrinsic activity of agonists and
suggest that Asn-293 is important for both properties of the agonist-receptor
interaction.
Wieland, K Zuurmond, H M Krasel, C Ijzerman, A P Lohse, M J Comparative
Study Research Support, Non-U.S. Gov't United states Proceedings
of the National Academy of Sciences of the United States of America
Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9276-81.
%0 Journal Article
%1 Wieland1996
%A Wieland, K.
%A Zuurmond, H. M.
%A Krasel, C.
%A Ijzerman, A. P.
%A Lohse, M. J.
%D 1996
%J Proc Natl Acad Sci U S A
%K Adenylate Adrenergic Alprenolol/metabolism Animals Asparagine/genetics/metabolism Binding CHO Computer Cricetinae Cyclase/drug Humans Isoproterenol/chemistry/*metabolism/pharmacology Metoprolol/metabolism Models, Molecular Mutagenesis, Mutation Propranolol/metabolism Serine/genetics/metabolism Signal Simulation Site-Directed Sites Stereoisomerism Transduction beta-2/agonists/genetics/*metabolism beta-Agonists/*metabolism effects Receptor Cell
%N 17
%P 9276-81
%T Involvement of Asn-293 in stereospecific agonist recognition and
in activation of the beta 2-adrenergic receptor
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8799191
%V 93
%X To investigate the molecular mechanism for stereospecific binding
of agonists to beta 2-adrenergic receptors we used receptor models
to identify potential binding sites for the beta-OH-group of the
ligand, which defines the chiral center. Ser-165, located in transmembrane
helix IV, and Asn-293, situated in the upper half of transmembrane
helix VI, were identified as potential binding sites. Mutation of
Ser-165 to Ala did not change the binding of either isoproterenol
isomer as revealed after transient expression in human embryonic
kidney (HEK)-293 cells. In contrast, a receptor mutant in which Asn-293
was replaced by Leu showed substantial loss of stereospecific isoproterenol
binding. Adenylyl cyclase stimulation by this mutant after stable
expression in CHO cells confirmed the substantial loss of stereospecificity
for isoproterenol. In a series of agonists the loss of affinity in
the Leu-293 mutant receptor was strongly correlated with the intrinsic
activity of the compounds. Full agonists showed a 10-30-fold affinity
loss, whereas partial agonists had almost the same affinity for both
receptors. Stereospecific recognition of antagonists was unaltered
in the Leu-293 mutant receptor. These data indicate a relationship
between stereospecificity and intrinsic activity of agonists and
suggest that Asn-293 is important for both properties of the agonist-receptor
interaction.
@article{Wieland1996,
abstract = {To investigate the molecular mechanism for stereospecific binding
of agonists to beta 2-adrenergic receptors we used receptor models
to identify potential binding sites for the beta-OH-group of the
ligand, which defines the chiral center. Ser-165, located in transmembrane
helix IV, and Asn-293, situated in the upper half of transmembrane
helix VI, were identified as potential binding sites. Mutation of
Ser-165 to Ala did not change the binding of either isoproterenol
isomer as revealed after transient expression in human embryonic
kidney (HEK)-293 cells. In contrast, a receptor mutant in which Asn-293
was replaced by Leu showed substantial loss of stereospecific isoproterenol
binding. Adenylyl cyclase stimulation by this mutant after stable
expression in CHO cells confirmed the substantial loss of stereospecificity
for isoproterenol. In a series of agonists the loss of affinity in
the Leu-293 mutant receptor was strongly correlated with the intrinsic
activity of the compounds. Full agonists showed a 10-30-fold affinity
loss, whereas partial agonists had almost the same affinity for both
receptors. Stereospecific recognition of antagonists was unaltered
in the Leu-293 mutant receptor. These data indicate a relationship
between stereospecificity and intrinsic activity of agonists and
suggest that Asn-293 is important for both properties of the agonist-receptor
interaction.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Wieland, K. and Zuurmond, H. M. and Krasel, C. and Ijzerman, A. P. and Lohse, M. J.},
biburl = {https://www.bibsonomy.org/bibtex/2589d1c7fd1b0bc1b4747dc13a5ce26ca/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {bf4c143133db8cc902c52e4bc6108525},
intrahash = {589d1c7fd1b0bc1b4747dc13a5ce26ca},
issn = {0027-8424 (Print) 0027-8424 (Linking)},
journal = {Proc Natl Acad Sci U S A},
keywords = {Adenylate Adrenergic Alprenolol/metabolism Animals Asparagine/genetics/metabolism Binding CHO Computer Cricetinae Cyclase/drug Humans Isoproterenol/chemistry/*metabolism/pharmacology Metoprolol/metabolism Models, Molecular Mutagenesis, Mutation Propranolol/metabolism Serine/genetics/metabolism Signal Simulation Site-Directed Sites Stereoisomerism Transduction beta-2/agonists/genetics/*metabolism beta-Agonists/*metabolism effects Receptor Cell},
month = {Aug 20},
note = {Wieland, K Zuurmond, H M Krasel, C Ijzerman, A P Lohse, M J Comparative
Study Research Support, Non-U.S. Gov't United states Proceedings
of the National Academy of Sciences of the United States of America
Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9276-81.},
number = 17,
pages = {9276-81},
shorttitle = {Involvement of Asn-293 in stereospecific agonist recognition and in
activation of the beta 2-adrenergic receptor},
timestamp = {2010-12-14T18:22:43.000+0100},
title = {Involvement of Asn-293 in stereospecific agonist recognition and
in activation of the beta 2-adrenergic receptor},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=8799191},
volume = 93,
year = 1996
}