TY - JOUR
T1 - Bioorthogonal click chemistry to assay mu-opioid receptor palmitoylation using 15-hexadecynoic acid and immunoprecipitation
AU - Ebersole (Anderson), Brittany
AU - Petko, Jessica A.
AU - Levenson, Robert
N1 - Funding Information:
We thank Bernhard Lüscher and Casey Kilpatrick (Penn State University, University Park) for suggesting the use of 15-HDYA. This study was supported by a grant from the National Institute on Drug Abuse (NIDA, DA025995) and a CURE grant to R.L. from the Pennsylvania Department of Health using Tobacco Settlement Funds. The funding agencies had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
PY - 2014/4/15
Y1 - 2014/4/15
N2 - We have developed a modification of bioorthogonal click chemistry to assay the palmitoylation of cellular proteins. This assay uses 15-hexadecynoic acid (15-HDYA) as a chemical probe in combination with protein immunoprecipitation using magnetic beads in order to detect S-palmitoylation of proteins of interest. Here we demonstrate the utility of this approach for the mu-opioid receptor (MOR), a G-protein-coupled receptor (GPCR) responsible for mediating the analgesic and addictive properties of most clinically relevant opioid agonist drugs. This technique provides a rapid, non-isotopic, and efficient method to assay the palmitoylation status of a variety of cellular proteins, including most GPCRs.
AB - We have developed a modification of bioorthogonal click chemistry to assay the palmitoylation of cellular proteins. This assay uses 15-hexadecynoic acid (15-HDYA) as a chemical probe in combination with protein immunoprecipitation using magnetic beads in order to detect S-palmitoylation of proteins of interest. Here we demonstrate the utility of this approach for the mu-opioid receptor (MOR), a G-protein-coupled receptor (GPCR) responsible for mediating the analgesic and addictive properties of most clinically relevant opioid agonist drugs. This technique provides a rapid, non-isotopic, and efficient method to assay the palmitoylation status of a variety of cellular proteins, including most GPCRs.
UR - http://www.scopus.com/inward/record.url?scp=84895547242&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84895547242&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2014.01.008
DO - 10.1016/j.ab.2014.01.008
M3 - Article
C2 - 24463015
AN - SCOPUS:84895547242
SN - 0003-2697
VL - 451
SP - 25
EP - 27
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -