Making structural sense of dimerization interfaces of delta opioid receptor homodimers

Jennifer M. Johnston, Mahalaxmi Aburi, Davide Provasi, Andrea Bortolato, Eneko Urizar, Nevin A Lambert, Jonathan A. Javitch, Marta Filizola

Research output: Contribution to journalArticle

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Abstract

Opioid receptors, like other members of the G protein-coupled receptor (GPCR) family, have been shown to associate to form dimers and/or oligomers at the plasma membrane. Whether this association is stable or transient is not known. Recent compelling evidence suggests that at least some GPCRs rapidly associate and dissociate. We have recently calculated binding affinities from free energy estimates to predict transient association between mouse delta opioid receptor (DOR) protomers at a symmetric interface involving the fourth transmembrane (TM4) helix (herein termed "4" dimer). Here we present disulfide cross-linking experiments with DOR constructs with cysteines substituted at the extracellular ends of TM4 or TM5 that confirm the formation of DOR complexes involving these helices. Our results are consistent with the involvement of TM4 and/or TM5 at the DOR homodimer interface, but possibly with differing association propensities. Coarse-grained (CG) well-tempered metadynamics simulations of two different dimeric arrangements of DOR involving TM4 alone or with TM5 (herein termed "4/5" dimer) in an explicit lipid-water environment confirmed the presence of two structurally and energetically similar configurations of the 4 dimer, as previously assessed by umbrella sampling calculations, and revealed a single energetic minimum of the 4/5 dimer. Additional CG umbrella sampling simulations of the 4/5 dimer indicated that the strength of association between DOR protomers varies depending on the protein region at the interface, with the 4 dimer being more stable than the 4/5 dimer.

Original languageEnglish (US)
Pages (from-to)1682-1690
Number of pages9
JournalBiochemistry
Volume50
Issue number10
DOIs
StatePublished - Mar 15 2011

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delta Opioid Receptor
Dimerization
Dimers
Protein Subunits
Association reactions
Opioid Receptors
G-Protein-Coupled Receptors
Disulfides
Cysteine
Sampling
Cell Membrane
Lipids
Cell membranes
Oligomers
Water
Free energy
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Johnston, J. M., Aburi, M., Provasi, D., Bortolato, A., Urizar, E., Lambert, N. A., ... Filizola, M. (2011). Making structural sense of dimerization interfaces of delta opioid receptor homodimers. Biochemistry, 50(10), 1682-1690. https://doi.org/10.1021/bi101474v

Making structural sense of dimerization interfaces of delta opioid receptor homodimers. / Johnston, Jennifer M.; Aburi, Mahalaxmi; Provasi, Davide; Bortolato, Andrea; Urizar, Eneko; Lambert, Nevin A; Javitch, Jonathan A.; Filizola, Marta.

In: Biochemistry, Vol. 50, No. 10, 15.03.2011, p. 1682-1690.

Research output: Contribution to journalArticle

Johnston, JM, Aburi, M, Provasi, D, Bortolato, A, Urizar, E, Lambert, NA, Javitch, JA & Filizola, M 2011, 'Making structural sense of dimerization interfaces of delta opioid receptor homodimers', Biochemistry, vol. 50, no. 10, pp. 1682-1690. https://doi.org/10.1021/bi101474v
Johnston JM, Aburi M, Provasi D, Bortolato A, Urizar E, Lambert NA et al. Making structural sense of dimerization interfaces of delta opioid receptor homodimers. Biochemistry. 2011 Mar 15;50(10):1682-1690. https://doi.org/10.1021/bi101474v
Johnston, Jennifer M. ; Aburi, Mahalaxmi ; Provasi, Davide ; Bortolato, Andrea ; Urizar, Eneko ; Lambert, Nevin A ; Javitch, Jonathan A. ; Filizola, Marta. / Making structural sense of dimerization interfaces of delta opioid receptor homodimers. In: Biochemistry. 2011 ; Vol. 50, No. 10. pp. 1682-1690.
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