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פותח על ידי קלירמאש פתרונות בע"מ -
Antibodies for the Immunochemistry of the Human β3‐Adrenergic Receptor
Year:
1994
Source of publication :
European Journal of Biochemistry
Authors :
אשדת, יובל
;
.
Volume :
224
Co-Authors:
Guillaume, J.‐L., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Petitjean, F., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Haasemann, M., Institut Jacques Monod, Université Paris VII, France
Bianchi, C., Center of Prevention of Cardiovascular Disease, Harvard School of Public Health, Boston, United States
Eshdat, Y., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Strosberg, A.D., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Facilitators :
From page:
761
To page:
770
(
Total pages:
10
)
Abstract:
Based on the amino acid sequence deduced from the recently cloned human β3‐adrenergic receptor (huβ3AR) gene, polyclonal antibodies were prepared against synthetic peptides, corresponding to regions of huβ3AR presumed to be exposed at the outer or the inner side of the membrane on the basis of the putative three‐dimensional structure of the previously characterized β1 and β2 adrenergic receptors. Affinity‐purified antibodies directed against N‐terminal, extracellular or intracellular loops and C‐terminal peptides reacted specifically with the huβ3AR and not with either the human β1 or β2 adrenergic receptor. Using these antibodies, it was demonstrated that the receptor is present at the surface of Chinese Hamster Ovary (CHO) cells transfected with the huβ3AR gene; in addition, the presence of the receptor protein was established in a human tissue (gall bladder). Immuno‐affinity chromatography of solubilized CHO huβ3AR‐containing cell membranes allowed the isolation of huβ3AR protein with an overall yield of 30%. The degree of purity of the receptor was more than 80%, as assessed by N‐terminal sequencing of the protein eluted from the column. Sequence analysis demonstrated the absence of a methionine residue at the N‐terminal position, and suggested that the side chain of the asparagine residue at position 7 is glycosylated. Copyright © 1994, Wiley Blackwell. All rights reserved
Note:
Related Files :
Animal
animal cell
Base Sequence
beta 3 adrenergic receptor
cell membrane
cho cell
Electrophoresis, Polyacrylamide Gel
peptides
עוד תגיות
תוכן קשור
More details
DOI :
10.1111/j.1432-1033.1994.00761.x
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
21683
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:46
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Scientific Publication
Antibodies for the Immunochemistry of the Human β3‐Adrenergic Receptor
224
Guillaume, J.‐L., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Petitjean, F., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Haasemann, M., Institut Jacques Monod, Université Paris VII, France
Bianchi, C., Center of Prevention of Cardiovascular Disease, Harvard School of Public Health, Boston, United States
Eshdat, Y., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Strosberg, A.D., Institut Cochin de Génétique Moléculaire, Laboratoire D'immuno-Pharmacologie Moléculaire, Centre National de la Recherche Scientifique Université, Paris, France
Antibodies for the Immunochemistry of the Human β3‐Adrenergic Receptor
Based on the amino acid sequence deduced from the recently cloned human β3‐adrenergic receptor (huβ3AR) gene, polyclonal antibodies were prepared against synthetic peptides, corresponding to regions of huβ3AR presumed to be exposed at the outer or the inner side of the membrane on the basis of the putative three‐dimensional structure of the previously characterized β1 and β2 adrenergic receptors. Affinity‐purified antibodies directed against N‐terminal, extracellular or intracellular loops and C‐terminal peptides reacted specifically with the huβ3AR and not with either the human β1 or β2 adrenergic receptor. Using these antibodies, it was demonstrated that the receptor is present at the surface of Chinese Hamster Ovary (CHO) cells transfected with the huβ3AR gene; in addition, the presence of the receptor protein was established in a human tissue (gall bladder). Immuno‐affinity chromatography of solubilized CHO huβ3AR‐containing cell membranes allowed the isolation of huβ3AR protein with an overall yield of 30%. The degree of purity of the receptor was more than 80%, as assessed by N‐terminal sequencing of the protein eluted from the column. Sequence analysis demonstrated the absence of a methionine residue at the N‐terminal position, and suggested that the side chain of the asparagine residue at position 7 is glycosylated. Copyright © 1994, Wiley Blackwell. All rights reserved
Scientific Publication
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