کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5813787 | 1556616 | 2015 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Engineering α4β2 nAChRs with reduced or increased nicotine sensitivity via selective disruption of consensus sites in the M3-M4 cytoplasmic loop of the α4 subunit
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کلمات کلیدی
HEPESCPMnAChRpKaCKIIPKCPKBN-(2-Hydroxyethyl)piperazine-N′-2-ethanesulfonic acid - N- (2-Hydroxyethyl) piperazine-N'-2-ethanesulfonic acidACh - آهAcetylcholine - استیل کولینTyrosine kinase - تیروزین کینازcounts per minute - شمار در هر دقیقهPhosphorylation - فسفریلاسیونNicotine - نیکوتین protein kinase A - پروتئین کیناز Aprotein kinase B - پروتئین کیناز BProtein kinase C - پروتئین کیناز سیcasein kinase II - کازئین کیناز IInicotinic acetylcholine receptor - گیرنده استیلکولین نیکوتینTwo-electrode voltage clamp - گیره ولتاژ دو الکترود
موضوعات مرتبط
علوم زیستی و بیوفناوری
علم عصب شناسی
علوم اعصاب رفتاری
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چکیده انگلیسی
The α4β2 neuronal nicotinic acetylcholine receptor (nAChR) plays a crucial role in nicotine addiction. These receptors are known to desensitize and up-regulate after chronic nicotine exposure, but the mechanism remains unknown. Currently, the structure and functional role of the intracellular domains of the nAChR are obscure. To study the effect of subunit phosphorylation on α4β2 nAChR function and expression, eleven residues located in the M3-M4 cytoplasmic loop were mutated to alanine and aspartic acid. Two-electrode voltage clamp and 125I-labeled epibatidine binding assays were performed on Xenopus oocytes to assess agonist activation and receptor expression. When ACh was used as an agonist, a decrease in receptor activation was observed for the majority of the mutations. When nicotine was used as an agonist, four mutations exhibited a statistically significant hypersensitivity to nicotine (S438D, S469A, Y576A, and S589A). Additionally, two mutations (S516D and T536A) that displayed normal activation with ACh displayed remarkable reductions in sensitivity to nicotine. Binding assays revealed a constitutive up-regulation in these two nicotine mutations with reduced nicotine sensitivity. These results suggest that consensus phosphorylation residues in the M3-M4 cytoplasmic loop of the α4 subunit play a crucial role in regulating α4β2 nAChR agonist selectivity and functional expression. Furthermore, these results suggest that disruption of specific interactions at PKC putative consensus sites can render α4β2 nAChRs almost insensitive to nicotine without substantial effects on normal AChR function. Therefore, these PKC consensus sites in the M3-M4 cytoplasmic loop of the α4 nAChR subunit could be a target for smoking cessation drugs.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Neuropharmacology - Volume 99, December 2015, Pages 273-284
Journal: Neuropharmacology - Volume 99, December 2015, Pages 273-284
نویسندگان
Nilza M. Biaggi-Labiosa, Emir Avilés-Pagán, Daniel Caballero-Rivera, Carlos A. Báez-Pagán, José A. Lasalde-Dominicci,