Agonist-mediated tyrosine phosphorylation of isoforms of the shc adapter protein by the delta opioid receptor.
Maximally effective concentrations of the opioid agonist D-ala2-D-leu5-enkephalin resulted in some 2-3-fold enhancement of tyrosine phosphorylation of the p52 Shc adapter protein in a clone of Rat-1 fibroblasts transfected to express stably the murine delta opioid receptor. More limited modifications of the tyrosine phosphorylation status of the p46 and p66 forms of Shc were observed in parallel. Epidermal growth factor caused some 10-12-fold enhancement of tyrosine phosphorylation of p52 Shc and marked increases in the p46 and p66 forms. The effect of D-ala2-D-leu5-enkephalin was prevented by pretreatment of the cells with pertussis toxin and was not observed in non-transfected parental fibroblasts whereas the effect of epidermal growth factor was still manifest in both these situations. Half-maximal effects of D-ala2-D-leu5-enkephalin on p52 Shc tyrosine phosphorylation were produced with sub-nanomolar concentrations, in agreement with previous results on the tyrosine phosphorylation of p44MAPK (Burt et al., 1996). p52 Shc became tyrosine phosphorylated more rapidly than p44MAPK in response to D-ala2-D-leu5-enkephalin and its enhanced tyrosine phosphorylation was maintained for at least 10 min.
Mullaney
I
I
Division of Biochemistry and Molecular Biology, University of Glasgow, Scotland, U.K.
Carr
I C
IC
Burt
A R
AR
Wilson
M
M
Anderson
N G
NG
Milligan
G
G
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D002999:Clone Cells / Q000737:chemistry / Q000201:enzymology; D016308:Enkephalin, Leucine-2-Alanine / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000378:metabolism / Q000494:pharmacology; D066246:ErbB Receptors / Q000819:agonists / Q000235:genetics; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D015870:Gene Expression / Q000502:physiology; D007536:Isomerism; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D037342:Pertussis Toxin; D010766:Phosphorylation; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017465:Receptors, Opioid, delta / Q000819:agonists* / Q000235:genetics; D055765:Shc Signaling Adaptor Proteins; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D013997:Time Factors; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D011506:Proteins; D017465:Receptors, Opioid, delta; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; C528035:Shc1 protein, rat; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D010566:Virulence Factors, Bordetella; D014443:Tyrosine; D004815:Epidermal Growth Factor; D016308:Enkephalin, Leucine-2-Alanine; D037342:Pertussis Toxin; D066246:ErbB Receptors; D019950:Mitogen-Activated Protein Kinase 1