Camphor modulates TRPV3 cation channels activity by interacting with critical pore-region cysteine residues
Pakistan Journal of Pharmaceutical Sciences. 2013; 26 (3): 431-438
Dans Anglais
| IMEMR
| ID: emr-142600
ABSTRACT
TRPV3 ion channels mediate thermo-transduction, nociception, inflammation and dermatitis in mammals. TRPV1-4 proteins have been shown to have conserved cysteine-residues in the pore-forming regions. These residues participate in channel activation via S-nitrosylation of channel proteins. Camphor is a commonly used ligand for TRPV3 channels. Thus the knowledge about the potential binding/interacting site[s] for camphor will help to design effective and potent analgesic compounds. In an overlap-extension PCR method, following primer-pairs were used to mutate conserved cysteine-residues in the pore-region of TRPV3 channels; GATTGAGAATcCTCCAAGGACAAAAAGGAC, TRPV3-C612S-Fw and GTCCTTGGAGgACTTCTCAATCAGTCAGTGAGG, TRPV3-C612S-Rv primers pair. And for TRPV3-C619S GGACTCcAGTTCCTATGGCCAGC, TRPV3-C619S-Fw and GCTGGCCATAgGAACTGGAGTCC, TRPV3-C619S-Rv respectively. All cDNA constructs were confirmed by DNA-sequencing and used to make cRNAs. Oocytes expressing mTRPV3-C619S and mTRPV3-C612S mutant channels were challenged with 2-APB [1 mM], camphor [10 mM] and dihydrocarveol [10 mM] either at -40 mV or +40 mV holding potentials in voltage-clamp experiments. Responses of both mutants to 2-APB were similar to wild-type mTRPV3. Interestingly, responses to camphor were totally lost in mTRPV3-C619S mutant, while responses to dihydrocarveol remained intact. In contrast mTRPV3-C612S displayed slightly altered [16 +/- 2% reduction] phenotype with respect to camphor sensitivity. It is concluded that pore-region cysteines play critical role in camphor sensitivity of TRPV3 ion channels
Recherche sur Google
Indice:
Méditerranée orientale
Sujet Principal:
Xenopus
/
Sites de fixation
/
Composés du bore
/
Camphre
/
Données de séquences moléculaires
/
Séquence d'acides aminés
/
ADN complémentaire
/
Cystéine
/
Monoterpènes
/
Souris
Type d'étude:
Guide de pratique
Limites du sujet:
Animaux
langue:
Anglais
Texte intégral:
Pak. J. Pharm. Sci.
Année:
2013
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