Facebook immune system

  • Stein T
  • Chen E
  • Mangla K
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The carboxyl tail of the human mu opioid receptor was shown to bind the carboxyl terminal region of human filamin A, a protein known to couple membrane proteins to actin. Results from yeast two-hybrid screening were confirmed by direct protein-protein binding and by coimmunoprecipitation of filamin and mu opioid receptor from cell lysates. To investigate the role of filamin A in opioid receptor function and regulation, we used the melanoma cell line M2, which does not express filamin A, and its subclone A7, transfected with human filamin A cDNA. Both cell lines were stably transfected with cDNA encoding myc-tagged human mu opioid receptor. Fluorescent studies, using confocal microscopy, provided evidence that filamin and mu opioid receptors were extensively colocalized on the membranes of filamin-expressing melanoma cells. The immunostaining of mu opioid receptors indicated that the lack of filamin had no detectable effect on membrane localization of the receptors. Moreover, mu opioid receptors function normally in the absence of filamin A, as evidenced by studies of opioid binding and DAMGO inhibition of forskolin-stimulated adenylyl cyclase. However, agonist-induced receptor down-regulation and functional desensitization were virtually abolished in cells lacking filamin A. The level of internalized mu-opioid receptors, after 30-min exposure to agonist, was greatly reduced, suggesting a role for filamin in mu opioid receptor trafficking. During these studies, we observed that forskolin activation of adenylyl cyclase was greatly reduced in filamin-lacking cells. An even more unexpected finding was the ability of long-term treatment with d-Ala2,N-Me-Phe4,Gly5-ol-enkephalin of M2 cells, containing mu opioid receptors, to restore normal forskolin activation. The mechanism of this effect is currently unknown. It is postulated that the observed effects on mu opioid receptor regulation by filamin A and, by implication, of the actin cytoskeleton may be the result of its role in mu opioid receptor trafficking.




Stein, T., Chen, E., & Mangla, K. (2011). Facebook immune system (pp. 1–8). Association for Computing Machinery (ACM). https://doi.org/10.1145/1989656.1989664

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