title: Mapping the binding site of TRPV1 on AKAP79: implications for inflammatory hyperalgesia.
abstract: Inflammation causes hyperalgesia, an enhanced sensitivity to noxious stimuli. Transient receptor potential vanilloid 1 (TRPV1), a thermo-TRP ion channel activated by painful levels of heat, is an important contributor because hyperalgesia is reduced when TRPV1 is either genetically deleted or pharmacologically blocked. Inflammatory mediators such as prostaglandin-E2 or bradykinin cause hyperalgesia by activating cellular kinases that phosphorylate TRPV1, a process that has recently been shown to rely on a scaffolding protein, AKAP79, to target the kinases to TRPV1. Here we use Förster resonance energy transfer, immunoprecipitation, and TRPV1 membrane trafficking experiments to identify a key region on AKAP79, between amino acids 326-336, which is responsible for its interaction with TRPV1. A peptide identical to this domain inhibited sensitization of TRPV1 in vitro, and when covalently linked to a TAT peptide to promote uptake across the cell membrane the peptide inhibited in vivo inflammatory hyperalgesia in mice. Critically, it did so without affecting pain thresholds in the absence of inflammation. These results suggest that antagonizing the TRPV1-AKAP79 interaction will be a useful strategy for inhibiting inflammatory hyperalgesia.
authors.affiliation: Department of Pharmacology, University of Cambridge, Cambridge CB2 1PD, United Kingdom.
authors.forename: Joan
authors.initials: J
authors.lastname: Btesh
authors.affiliation: Department of Pharmacology, University of Cambridge, Cambridge CB2 1PD, United Kingdom.|Institute of Physiology and Pathophysiology, University of Erlangen-Nuremberg, 91052 Erlangen, Germany.
authors.forename: Michael J M
authors.initials: MJM
authors.lastname: Fischer
authors.affiliation: Department of Biochemistry, University of Cambridge, Cambridge CB2 1 GA, United Kingdom.
authors.forename: Katherine
authors.initials: K
authors.lastname: Stott
authors.affiliation: Department of Pharmacology, University of Cambridge, Cambridge CB2 1PD, United Kingdom.
authors.forename: Peter A
authors.initials: PA
authors.lastname: McNaughton
mesh_terms: D054758:A Kinase Anchor Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000187:drug effects / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D002211:Capsaicin / Q000494:pharmacology; D002351:Carrageenan / Q000633:toxicity; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D006930:Hyperalgesia / Q000139:chemically induced / Q000188:drug therapy / Q000378:metabolism*; D047468:Immunoprecipitation; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation / Q000235:genetics; D009474:Neurons; D017288:Pain Threshold / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000627:therapeutic use; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D054758:A Kinase Anchor Proteins; C487775:AKAP5 protein, human; D001426:Bacterial Proteins; D008164:Luminescent Proteins; D010455:Peptides; D050916:TRPV Cation Channels; C494482:TRPV1 protein, human; C054562:yellow fluorescent protein, Bacteria; D002351:Carrageenan; D002211:Capsaicin; D002118:Calcium