We propose to characterize structurally and functionally the mu type opioid receptor, which we have recently purified to homogeneity in our laboratory. Specifically, we will: (1) reconstitute the receptor into a membrane environment, and attempt to demonstrate restoration of both opioid binding and opioid-receptor mediated function; (2) raise monoclonal and polyclonal antibodies to the receptor, and use them to (a) map the receptor's distribution in the brain; (b) determine the role of different portions of the receptor on binding and function; (c) construct an immunoaffinity column for purifying large quantities of the receptor; and (d) clone the receptor (see also aim #4, below); (3) to run several critical tests of receptor models and to study the molecular mechanisms of opiate binding to its receptor, including (a) determining whether negative cooperativity occurs during opiate binding; (b) use thermodynamic analysis of opiate binding, to determine the molecular mechanism of agoinst and antagonist interactions; (c) determining the regulatory effects of ions, GTP, and lipids on opioid-receptor interactions; (d) determining whether the mu type opioid receptor can inter- convert to other types, such as delta or kappa; and (e) determining by physical methods whether receptor conformational changes occur during opiate binding; and (4) clone the opiate receptor, by synthesis of oligodeoxynucleotide probes corresponding to the N- terminal sequence of the receptor, use of these to isolate opioid receptor mRNA, and insertion of the corresponding DNA into a cloning vector. In addition, the availability of receptor antibody should provide an alternative means for receptor cloning (see specific aim #2, above). These studies will make it possible to clarify the role opioid receptors play in analgesia and other clinically important effects, and to determine the biochemical processes by which these effects are produced.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
5R01DA001583-22
Application #
2897669
Study Section
Special Emphasis Panel (ZDA1-MXC-A (05))
Program Officer
Koustova, Elena
Project Start
1988-12-01
Project End
2002-03-31
Budget Start
1999-04-15
Budget End
2000-03-31
Support Year
22
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Pharmacology
Type
Schools of Medicine
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Hwang, Cheol Kyu; Wagley, Yadav; Law, Ping-Yee et al. (2017) Phosphorylation of poly(rC) binding protein 1 (PCBP1) contributes to stabilization of mu opioid receptor (MOR) mRNA via interaction with AU-rich element RNA-binding protein 1 (AUF1) and poly A binding protein (PABP). Gene 598:113-130
Song, Kyu Young; Choi, Hack Sun; Law, Ping-Yee et al. (2017) Post-Transcriptional Regulation of the Human Mu-Opioid Receptor (MOR) by Morphine-Induced RNA Binding Proteins hnRNP K and PCBP1. J Cell Physiol 232:576-584
Wagley, Yadav; Law, Ping-Yee; Wei, Li-Na et al. (2017) Epigenetic Activation of ?-Opioid Receptor Gene via Increased Expression and Function of Mitogen- and Stress-Activated Protein Kinase 1. Mol Pharmacol 91:357-372
Hwang, Cheol Kyu; Wagley, Yadav; Law, Ping-Yee et al. (2015) Analysis of epigenetic mechanisms regulating opioid receptor gene transcription. Methods Mol Biol 1230:39-51
Wei, Li-Na; Dmintrovsky, Ethan (2015) Retinoids are back. FASEB J 29:1131-5
Macias Konstantopoulos, Wendy L; Dreifuss, Jessica A; McDermott, Katherine A et al. (2014) Identifying patients with problematic drug use in the emergency department: results of a multisite study. Ann Emerg Med 64:516-25
Feng, X; Wu, C-Y; Burton, F H et al. (2014) ?-arrestin protects neurons by mediating endogenous opioid arrest of inflammatory microglia. Cell Death Differ 21:397-406
Wagley, Yadav; Hwang, Cheol Kyu; Lin, Hong-Yiou et al. (2013) Inhibition of c-Jun NH2-terminal kinase stimulates mu opioid receptor expression via p38 MAPK-mediated nuclear NF-?B activation in neuronal and non-neuronal cells. Biochim Biophys Acta 1833:1476-88
Wu, Qifang; Hwang, Cheol Kyu; Zheng, Hui et al. (2013) MicroRNA 339 down-regulates ?-opioid receptor at the post-transcriptional level in response to opioid treatment. FASEB J 27:522-35
Song, Kyu Young; Choi, Hack Sun; Law, Ping-Yee et al. (2013) Vimentin interacts with the 5'-untranslated region of mouse mu opioid receptor (MOR) and is required for post-transcriptional regulation. RNA Biol 10:256-66

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