Methods to detect cell surface expression and constitutive activity of GPR6

Balakrishna M. Prasad, Bettye Hollins, Nevin A Lambert

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Citations (Scopus)

Abstract

GPR6 is a constitutively active Gs-coupled receptor that can signal from intracellular compartments. We present different methods used to study cell surface expression of receptors and other membrane proteins. A comparison of these methods shows that methods based on susceptibility to proteolytic enzymes are more efficient at providing estimates of cell surface expression than the commonly used cell surface biotinylation method. We also present different methods that can be used to detect constitutive activity of Gs-coupled receptors. Imaging-based assays to detect intracellular cyclic AMP accumulation are well suited to study signaling at a single cell level. These assays are particularly useful when the cells of interest form a small fraction of the culture such as primary cultures with low transfection efficiency.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages179-195
Number of pages17
EditionC
DOIs
StatePublished - Jan 1 2010

Publication series

NameMethods in Enzymology
NumberC
Volume484
ISSN (Print)0076-6879

Fingerprint

Assays
Cyclic AMP
Membrane Proteins
Biotinylation
Peptide Hydrolases
Imaging techniques
Cell Surface Receptors
Transfection

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Prasad, B. M., Hollins, B., & Lambert, N. A. (2010). Methods to detect cell surface expression and constitutive activity of GPR6. In Methods in Enzymology (C ed., pp. 179-195). (Methods in Enzymology; Vol. 484, No. C). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-381298-8.00010-1

Methods to detect cell surface expression and constitutive activity of GPR6. / Prasad, Balakrishna M.; Hollins, Bettye; Lambert, Nevin A.

Methods in Enzymology. C. ed. Academic Press Inc., 2010. p. 179-195 (Methods in Enzymology; Vol. 484, No. C).

Research output: Chapter in Book/Report/Conference proceedingChapter

Prasad, BM, Hollins, B & Lambert, NA 2010, Methods to detect cell surface expression and constitutive activity of GPR6. in Methods in Enzymology. C edn, Methods in Enzymology, no. C, vol. 484, Academic Press Inc., pp. 179-195. https://doi.org/10.1016/B978-0-12-381298-8.00010-1
Prasad BM, Hollins B, Lambert NA. Methods to detect cell surface expression and constitutive activity of GPR6. In Methods in Enzymology. C ed. Academic Press Inc. 2010. p. 179-195. (Methods in Enzymology; C). https://doi.org/10.1016/B978-0-12-381298-8.00010-1
Prasad, Balakrishna M. ; Hollins, Bettye ; Lambert, Nevin A. / Methods to detect cell surface expression and constitutive activity of GPR6. Methods in Enzymology. C. ed. Academic Press Inc., 2010. pp. 179-195 (Methods in Enzymology; C).
@inbook{b6dde33473b04d4fb0528055d561f62f,
title = "Methods to detect cell surface expression and constitutive activity of GPR6",
abstract = "GPR6 is a constitutively active Gs-coupled receptor that can signal from intracellular compartments. We present different methods used to study cell surface expression of receptors and other membrane proteins. A comparison of these methods shows that methods based on susceptibility to proteolytic enzymes are more efficient at providing estimates of cell surface expression than the commonly used cell surface biotinylation method. We also present different methods that can be used to detect constitutive activity of Gs-coupled receptors. Imaging-based assays to detect intracellular cyclic AMP accumulation are well suited to study signaling at a single cell level. These assays are particularly useful when the cells of interest form a small fraction of the culture such as primary cultures with low transfection efficiency.",
author = "Prasad, {Balakrishna M.} and Bettye Hollins and Lambert, {Nevin A}",
year = "2010",
month = "1",
day = "1",
doi = "10.1016/B978-0-12-381298-8.00010-1",
language = "English (US)",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
number = "C",
pages = "179--195",
booktitle = "Methods in Enzymology",
address = "United States",
edition = "C",

}

TY - CHAP

T1 - Methods to detect cell surface expression and constitutive activity of GPR6

AU - Prasad, Balakrishna M.

AU - Hollins, Bettye

AU - Lambert, Nevin A

PY - 2010/1/1

Y1 - 2010/1/1

N2 - GPR6 is a constitutively active Gs-coupled receptor that can signal from intracellular compartments. We present different methods used to study cell surface expression of receptors and other membrane proteins. A comparison of these methods shows that methods based on susceptibility to proteolytic enzymes are more efficient at providing estimates of cell surface expression than the commonly used cell surface biotinylation method. We also present different methods that can be used to detect constitutive activity of Gs-coupled receptors. Imaging-based assays to detect intracellular cyclic AMP accumulation are well suited to study signaling at a single cell level. These assays are particularly useful when the cells of interest form a small fraction of the culture such as primary cultures with low transfection efficiency.

AB - GPR6 is a constitutively active Gs-coupled receptor that can signal from intracellular compartments. We present different methods used to study cell surface expression of receptors and other membrane proteins. A comparison of these methods shows that methods based on susceptibility to proteolytic enzymes are more efficient at providing estimates of cell surface expression than the commonly used cell surface biotinylation method. We also present different methods that can be used to detect constitutive activity of Gs-coupled receptors. Imaging-based assays to detect intracellular cyclic AMP accumulation are well suited to study signaling at a single cell level. These assays are particularly useful when the cells of interest form a small fraction of the culture such as primary cultures with low transfection efficiency.

UR - http://www.scopus.com/inward/record.url?scp=78049243428&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78049243428&partnerID=8YFLogxK

U2 - 10.1016/B978-0-12-381298-8.00010-1

DO - 10.1016/B978-0-12-381298-8.00010-1

M3 - Chapter

C2 - 21036233

AN - SCOPUS:78049243428

T3 - Methods in Enzymology

SP - 179

EP - 195

BT - Methods in Enzymology

PB - Academic Press Inc.

ER -