PMID- 10215864
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20190620
IS  - 0014-2956 (Print)
IS  - 0014-2956 (Linking)
VI  - 261
IP  - 2
DP  - 1999 Apr
TI  - Nonselective coupling of the human mu-opioid receptor to multiple inhibitory
      G-protein isoforms.
PG  - 517-23
AB  - The human mu-opioid receptor was expressed in Saccharomyces cerevisiae. Binding
      of [3H]diprenorphine to yeast spheroplasts was specific and saturable (Kd = 1 nm,
      Bmax = 0.2-1 pmol x mg-1 of membrane proteins). Inhibition of [3H]diprenorphine
      binding by antagonists and agonists with varying opioid selectivities (mu, delta 
      and kappa) occurred with the same order of potency as in mammalian tissues.
      Affinities of antagonists were the same with yeast spheroplasts as in reference
      tissues whereas those of agonists, except etorphine and buprenorphine, were
      10-fold to 100-fold lower. Addition of heterotrimeric Gi,o-proteins purified from
      bovine brain shifted the mu-opioid receptor into a high-affinity state for
      agonists. Using individually purified Galpha-subunits re-associated with
      betagamma-dimers, we showed that alphao1, alphao2, alphai1, alphai2 and alphai3
      reconstituted high-affinity agonist binding with equal efficiency. This suggests 
      that the structural determinants of the mu-opioid receptor responsible for
      G-protein coupling are not able to confer a high degree of specificity towards
      any member of the Gi,o family. The selective effects of opioid observed in
      specialized tissues upon opioid stimulation may be a result of regulation of
      G-protein activity by cell-specific factors which should conveniently be analysed
      using the reconstitution assay described here.
FAU - Gaibelet, G
AU  - Gaibelet G
AD  - CNRS (Unite propre 9062), Institut National des Sciences Appliquees, Institut de 
      Pharmacologie et de Biologie Structurale, Toulouse, France.
FAU - Meilhoc, E
AU  - Meilhoc E
FAU - Riond, J
AU  - Riond J
FAU - Saves, I
AU  - Saves I
FAU - Exner, T
AU  - Exner T
FAU - Liaubet, L
AU  - Liaubet L
FAU - Nurnberg, B
AU  - Nurnberg B
FAU - Masson, J M
AU  - Masson JM
FAU - Emorine, L J
AU  - Emorine LJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Eur J Biochem
JT  - European journal of biochemistry
JID - 0107600
RN  - 0 (Enkephalins)
RN  - 0 (Protein Isoforms)
RN  - 0 (Receptors, Opioid, mu)
RN  - 0 (Recombinant Proteins)
RN  - 100929-53-1 (Enkephalin, Ala(2)-MePhe(4)-Gly(5)-)
RN  - 1F0L5N25ZZ (Diprenorphine)
RN  - 34273-04-6 (Guanylyl Imidodiphosphate)
RN  - 40D3SCR4GZ (Buprenorphine)
RN  - 42M2Y6NU9O (Etorphine)
RN  - EC 3.6.1.- (GTP-Binding Proteins)
SB  - IM
MH  - Binding Sites
MH  - Binding, Competitive
MH  - Buprenorphine/metabolism
MH  - Diprenorphine/metabolism
MH  - Enkephalin, Ala(2)-MePhe(4)-Gly(5)-
MH  - Enkephalins/metabolism
MH  - Etorphine/metabolism
MH  - GTP-Binding Proteins/*metabolism
MH  - Gene Expression/genetics
MH  - Guanylyl Imidodiphosphate/metabolism
MH  - Humans
MH  - Protein Binding
MH  - Protein Isoforms/genetics
MH  - Receptors, Opioid, mu/*agonists/*antagonists & inhibitors
MH  - Recombinant Proteins/genetics
MH  - Saccharomyces cerevisiae/genetics
MH  - Spheroplasts/metabolism
MH  - Transformation, Genetic
EDAT- 1999/04/24 00:00
MHDA- 1999/04/24 00:01
CRDT- 1999/04/24 00:00
PHST- 1999/04/24 00:00 [pubmed]
PHST- 1999/04/24 00:01 [medline]
PHST- 1999/04/24 00:00 [entrez]
AID - 10.1046/j.1432-1327.1999.00301.x [doi]
PST - ppublish
SO  - Eur J Biochem. 1999 Apr;261(2):517-23. doi: 10.1046/j.1432-1327.1999.00301.x.