Effects of kyotorphin (L-tyrosyl-L-arginine) ON[3H]NG-nitro-L-arginine binding to neuronal nitric oxide synthase in rat brain.
L-Tyrosyl-L-arginine (kyotorphin) is known as an endogenous analgesic neuropeptide. We examined whether kyotorphin and other arginine-containing neuropeptides were endogenous substrates for neuronal nitric oxide synthase (NOS) in the rat brain. Cytosol fractions of the rat cerebellum contained higher concentrations of neuronal NOS (nNOS) than endothelial NOS. In rat cerebellar cytosol, the binding activity of [3H]NG-nitro-L-arginine (NNA) was inhibited equally by L-arginine (L-Arg), kyotorphin, and L-leucyl-L-Arg (a kyotorphin receptor antagonist). Binding activities were inhibited to lesser degrees by fibronectin active fragments, bradykinin, and dynorphin A, but were not inhibited by L-tyrosyl-D-Arg or substance P. Interestingly, the inhibition of [3H]NNA binding by kyotorphin was attenuated by inhibitors of kyotorphin-hydrolyzing peptidases (KTPases) such as bestatin and arphamenine B. These results suggest that kyotorphin is degraded to L-Arg by KTPases, which in turn may act as substrate for nNOS.
Arima
T
T
Department of Pharmacology, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Kitamura
Y
Y
Nishiya
T
T
Taniguchi
T
T
Takagi
H
H
Nomura
Y
Y
D000818:Animals; D001120:Arginine / Q000494:pharmacology; D001920:Bradykinin / Q000494:pharmacology; D001921:Brain / Q000201:enzymology*; D002531:Cerebellum / Q000201:enzymology; D002540:Cerebral Cortex / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004151:Dipeptides / Q000494:pharmacology; D004399:Dynorphins / Q000494:pharmacology; D004723:Endorphins / Q000494:pharmacology*; D005353:Fibronectins / Q000494:pharmacology; D006624:Hippocampus / Q000201:enzymology; D015151:Immunoblotting; D009474:Neurons / Q000201:enzymology*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D019335:Nitroarginine / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014316:Tritium
D004151:Dipeptides; D004723:Endorphins; D005353:Fibronectins; C019448:kyotorphin; D014316:Tritium; D019335:Nitroarginine; C078789:leucylarginine; D004399:Dynorphins; D001120:Arginine; D019001:Nitric Oxide Synthase; D001920:Bradykinin