The interaction between the mu opioid receptor and filamin A.
Our laboratory embarked on research to discover proteins the interaction of which with the mu opioid receptor (MOPr) is required for its function and regulation. We performed yeast two-hybrid screens, using the carboxy tail of the human MOPr as bait and a human brain library. This yielded a number of proteins that seemed to bind to the MOPr C-tail. The one we chose to study in detail was filamin A (FLNA). Evidence was obtained that there was indeed protein-protein binding between the C-tail of MOPr and FLNA. A human melanoma cell line (M2) lacking the gene for FLNA and a control cell line (A7) which differed from M2 only in having been transfected with the gene for FLNA and expressing the FLNA protein were made available to us. We transfected these cell lines with the gene for MOPr and used them in our studies. The absence of FLNA strongly reduced MOPr downregulation as well as desensitization of adenylyl cyclase inhibition and G protein activation. A recent finding, published here for the first time, is that FLNA is required for the activation by mu opioid agonists of the MAP kinase p38. Deletion studies indicated that the MOPr binding site on FLNA is in the 24th repeat, close to its C-terminal. It was further found that FLNA lacking the N-terminal actin binding domain is as capable as full length FLNA to restore cells to control status, suggesting that actin binding is not required. A surprising finding was that upregulation of MOPr by morphine and some agonist analogs occurs in M2 cells lacking FLNA, whereas normal receptor downregulation takes place in A7 cells.
Simon
Eric J
EJ
Departments of Psychiatry and Pharmacology, NYU School of Medicine, 550 First Avenue, New York, NY 10016, USA. eric.simon@nyu.edu
Onoprishvili
Irma
I
D045744:Cell Line, Tumor; D003285:Contractile Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D064448:Filamins; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000473:pathology; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009020:Morphine / Q000008:administration & dosage / Q000494:pharmacology; D011485:Protein Binding; D017450:Receptors, Opioid, mu / Q000378:metabolism*; D017384:Sequence Deletion; D020798:Two-Hybrid System Techniques; D015854:Up-Regulation / Q000187:drug effects
D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D017450:Receptors, Opioid, mu; D009020:Morphine