Abstract
Dopamine receptors are a class of metabotropic G protein-coupled receptors. Plasma membrane expression is a key determinant of receptor signaling, and one that is regulated both by extra and intracellular cues. Abnormal dopamine receptor signaling is implicated in several neuropsychiatric disorders, including schizophrenia and attention deficit hyperactivity disorder, as well as drug abuse. Here, we describe in detail the application of two complementary applications of protein biotinylation and enzyme-linked immunoabsorbent assay (ELISA) for detecting and quantifying levels of dopamine receptors expressed on the cell surface. In the biotinylation method, cell surface receptors are labeled with Sulfo-NHS-biotin. The charge on the sulfonyl facilitates water solubility of the reactive biotin compound and prevents its diffusion across the plasma membrane. In the ELISA method, surface labeling is achieved with antibodies specific to extracellular epitopes on the receptors, and by fixing the cells without detergent such that the plasma membrane remains intact.
| Original language | English (US) |
|---|---|
| Title of host publication | Dopamine |
| Subtitle of host publication | Methods and Protocols |
| Publisher | Humana Press Inc. |
| Pages | 3-13 |
| Number of pages | 11 |
| ISBN (Print) | 9781627032506 |
| DOIs | |
| State | Published - 2013 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 964 |
| ISSN (Print) | 1064-3745 |
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Keywords
- ADHD
- Biotinylation
- DAPI
- ELISA
- Plasma membrane
- Schizophrenia
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Detection of cell surface dopamine receptors. / Xiao, Jiping; Bergson, Clare.
Dopamine: Methods and Protocols. Humana Press Inc., 2013. p. 3-13 (Methods in Molecular Biology; Vol. 964).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Detection of cell surface dopamine receptors
AU - Xiao, Jiping
AU - Bergson, Clare
PY - 2013
Y1 - 2013
N2 - Dopamine receptors are a class of metabotropic G protein-coupled receptors. Plasma membrane expression is a key determinant of receptor signaling, and one that is regulated both by extra and intracellular cues. Abnormal dopamine receptor signaling is implicated in several neuropsychiatric disorders, including schizophrenia and attention deficit hyperactivity disorder, as well as drug abuse. Here, we describe in detail the application of two complementary applications of protein biotinylation and enzyme-linked immunoabsorbent assay (ELISA) for detecting and quantifying levels of dopamine receptors expressed on the cell surface. In the biotinylation method, cell surface receptors are labeled with Sulfo-NHS-biotin. The charge on the sulfonyl facilitates water solubility of the reactive biotin compound and prevents its diffusion across the plasma membrane. In the ELISA method, surface labeling is achieved with antibodies specific to extracellular epitopes on the receptors, and by fixing the cells without detergent such that the plasma membrane remains intact.
AB - Dopamine receptors are a class of metabotropic G protein-coupled receptors. Plasma membrane expression is a key determinant of receptor signaling, and one that is regulated both by extra and intracellular cues. Abnormal dopamine receptor signaling is implicated in several neuropsychiatric disorders, including schizophrenia and attention deficit hyperactivity disorder, as well as drug abuse. Here, we describe in detail the application of two complementary applications of protein biotinylation and enzyme-linked immunoabsorbent assay (ELISA) for detecting and quantifying levels of dopamine receptors expressed on the cell surface. In the biotinylation method, cell surface receptors are labeled with Sulfo-NHS-biotin. The charge on the sulfonyl facilitates water solubility of the reactive biotin compound and prevents its diffusion across the plasma membrane. In the ELISA method, surface labeling is achieved with antibodies specific to extracellular epitopes on the receptors, and by fixing the cells without detergent such that the plasma membrane remains intact.
KW - ADHD
KW - Biotinylation
KW - DAPI
KW - ELISA
KW - Plasma membrane
KW - Schizophrenia
UR - http://www.scopus.com/inward/record.url?scp=84878630110&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84878630110&partnerID=8YFLogxK
U2 - 10.1007/978-1-62703-251-3_1
DO - 10.1007/978-1-62703-251-3_1
M3 - Chapter
C2 - 23296774
AN - SCOPUS:84878630110
SN - 9781627032506
T3 - Methods in Molecular Biology
SP - 3
EP - 13
BT - Dopamine
PB - Humana Press Inc.
ER -