Effect of synthetic cell-penetrating peptides on TrkA activity in PC12 cells.
As TrkA, a high-affinity receptor of nerve growth factor (NGF), is a potential target for relieving uncontrolled inflammatory pain, an effective inhibitor of TrkA has been required for pain management. To identify a specific inhibitor of TrkA activity, we designed cell-penetrating peptides combined with amino-acid sequences in the activation loop of TrkA to antagonize tyrosine kinase activity. To select a peptide inhibiting TrkA activity, we examined the effect of cell-penetrating peptides on tyrosine kinase activity of recombinant TrkA in vitro and studied their effects on NGF-stimulated neurite outgrowth and protein phosphorylation in PC12 cells. Thereafter we investigated the effect of the selected peptide on NGF-stimulated TrkA activity and the expression of transient receptor potential channel 1 in PC12 cells. The selected peptide inhibited TrkA activity, but did not inhibit tyrosine kinase activities of other receptor-type tyrosine kinases in vitro. It also suppressed NGF-stimulated responses in PC12 cells. The selected synthetic cell-penetrating peptide antagonizing TrkA function would be a candidate for inflammatory pain therapy.
Hirose
Munetaka
M
Department of Anesthesiology and Reanimatology, Faculty of Medical Sciences, Fukui University, Eiheijicho, Yoshidagun, Fukui, Japan. hirosem@u-fukui.ac.jp
Takatori
Mayumi
M
Kuroda
Yoshihiro
Y
Abe
Mineo
M
Murata
Eri
E
Isada
Tetsuro
T
Ueda
Koyo
K
Shigemi
Kenji
K
Shibazaki
Masayuki
M
Shimizu
Fumihiro
F
Hirata
Masashi
M
Fukazawa
Keita
K
Sakaguchi
Masahiro
M
Kageyama
Kyoko
K
Tanaka
Yoshifumi
Y
D000700:Analgesics / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002463:Cell Membrane Permeability; D004305:Dose-Response Relationship, Drug; D020932:Nerve Growth Factor / Q000378:metabolism; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism / Q000494:pharmacology*; D016716:PC12 Cells; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology*; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D051381:Rats; D020917:Receptor, trkA / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D050052:TRPC Cation Channels / Q000187:drug effects / Q000378:metabolism
D000700:Analgesics; C551391:IPTRK3 peptide; D009842:Oligopeptides; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; D050052:TRPC Cation Channels; C096351:transient receptor potential cation channel, subfamily C, member 1; D020932:Nerve Growth Factor; D020917:Receptor, trkA