A member of the heat shock protein 40 family, hlj1, binds to the carboxyl tail of the human mu opioid receptor.
A yeast two-hybrid screen, using the carboxyl tail of the human mu opioid receptor as bait and a human brain cDNA library as target, indicated that the carboxyl terminal portion of hlj1, a member of the human heat shock protein 40 family, interacts with the carboxyl tail of the human mu opioid receptor. To determine if direct in vitro binding occurs between these two proteins, we performed overlay experiments. Results from the overlay experiments showed that binding occurs between the His fusion protein of hlj1 and the GST fusion protein of the carboxyl tail of the human mu opioid receptor. In contrast, no binding with the His fusion protein of hlj1 occurred with GST alone or the GST fusion protein of the third cytoplasmic loop of the human mu opioid receptor. Results from co-immunoprecipitation studies, carried out in whole HEK cell lysates, confirmed in vivo binding between these two proteins. Immunofluorescent studies, using laser scanning confocal microscopy, showed significant co-localization between hlj1 and the human mu opioid receptor in the cell membrane. The function of this protein-protein interaction and its physiological relevance in animal and human brain is yet to be determined.
Department of Psychiatry, New York University School of Medicine, 550 First Avenue, New York, NY 10016, USA.
N
N
Ancevska-Taneva
I
I
Onoprishvili
M L
ML
Andria
J M
JM
Hiller
E J
EJ
Simon
D001665:Binding Sites / Q000502:physiology; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D050956:HSP40 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D047468:Immunoprecipitation / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D016296:Mutagenesis / Q000502:physiology; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D017450:Receptors, Opioid, mu / Q000737:chemistry / Q000378:metabolism*; D014162:Transfection / Q000379:methods; D020798:Two-Hybrid System Techniques
C494650:DNAJB4 protein, human; D050956:HSP40 Heat-Shock Proteins; D017450:Receptors, Opioid, mu