MOR Is Not Enough: Identification of Novel mu-Opioid Receptor Interacting Proteins Using Traditional and Modified Membrane Yeast Two-Hybrid Screens

Jessica A. Petko, Stephanie Justice-Bitner, Jay Jin, Victoria Wong, Saranya Kittanakom, Thomas N. Ferraro, Igor Stagljar, Robert Levenson

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The mu-opioid receptor (MOR) is the G-protein coupled receptor primarily responsible for mediating the analgesic and rewarding properties of opioid agonist drugs such as morphine, fentanyl, and heroin. We have utilized a combination of traditional and modified membrane yeast two-hybrid screening methods to identify a cohort of novel MOR interacting proteins (MORIPs). The interaction between the MOR and a subset of MORIPs was validated in pulldown, co-immunoprecipitation, and co-localization studies using HEK293 cells stably expressing the MOR as well as rodent brain. Additionally, a subset of MORIPs was found capable of interaction with the delta and kappa opioid receptors, suggesting that they may represent general opioid receptor interacting proteins (ORIPS). Expression of several MORIPs was altered in specific mouse brain regions after chronic treatment with morphine, suggesting that these proteins may play a role in response to opioid agonist drugs. Based on the known function of these newly identified MORIPs, the interactions forming the MOR signalplex are hypothesized to be important for MOR signaling and intracellular trafficking. Understanding the molecular complexity of MOR/MORIP interactions provides a conceptual framework for defining the cellular mechanisms of MOR signaling in brain and may be critical for determining the physiological basis of opioid tolerance and addiction.

Original languageEnglish (US)
Article numbere67608
JournalPloS one
Volume8
Issue number6
DOIs
StatePublished - Jun 28 2013

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Receptor-Interacting Protein Serine-Threonine Kinases
mu Opioid Receptor
narcotics
Yeast
Yeasts
yeasts
Membranes
receptors
proteins
Opioid Analgesics
Brain
Proteins
Morphine
morphine
kappa Opioid Receptor
Two-Hybrid System Techniques
delta Opioid Receptor
brain
HEK293 Cells
agonists

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Petko, Jessica A. ; Justice-Bitner, Stephanie ; Jin, Jay ; Wong, Victoria ; Kittanakom, Saranya ; Ferraro, Thomas N. ; Stagljar, Igor ; Levenson, Robert. / MOR Is Not Enough : Identification of Novel mu-Opioid Receptor Interacting Proteins Using Traditional and Modified Membrane Yeast Two-Hybrid Screens. In: PloS one. 2013 ; Vol. 8, No. 6.
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MOR Is Not Enough : Identification of Novel mu-Opioid Receptor Interacting Proteins Using Traditional and Modified Membrane Yeast Two-Hybrid Screens. / Petko, Jessica A.; Justice-Bitner, Stephanie; Jin, Jay; Wong, Victoria; Kittanakom, Saranya; Ferraro, Thomas N.; Stagljar, Igor; Levenson, Robert.

In: PloS one, Vol. 8, No. 6, e67608, 28.06.2013.

Research output: Contribution to journalArticle

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