کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5813764 | 1556616 | 2015 | 16 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Flexible subunit stoichiometry of functional human P2X2/3 heteromeric receptors
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کلمات کلیدی
BN-PAGEtrinitrophenyl-ATP2-methylthio ATPTNP-ATPP2X2/3 receptorsSubunit stoichiometryImaxLGICASIC2-MeSATPDRGEC50GFPdorsal root ganglion - گانگلیون ریشه پشتیBlue native PAGE - PAGE آبی بومیα,β-MeATP - α، β-MeATPα,β-methylene ATP - α، β-methylene ATPHill coefficient - ضریب هیلMutagenesis - موتاژنزwild type - نوع وحشیhalf maximal effective concentration - نیمه حداکثر غلظت موثرgreen fluorescent protein - پروتئین فلورسنت سبزAcid-sensing ion channel - کانال یون حساس به اسیدligand-gated ion channel - کانال یون لیگاند دار
موضوعات مرتبط
علوم زیستی و بیوفناوری
علم عصب شناسی
علوم اعصاب رفتاری
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The aim of the present work was to clarify whether heterotrimeric P2X2/3 receptors have a fixed subunit stoichiometry consisting of one P2X2 and two P2X3 subunits as previously suggested, or a flexible stoichiometry containing also the inverse subunit composition. For this purpose we transfected HEK293 cells with P2X2 and P2X3 encoding cDNA at the ratios of 1:2 and 4:1, and analysed the biophysical and pharmacological properties of the generated receptors by means of the whole-cell patch-clamp technique. The concentration-response curves for the selective agonist α,β-meATP did not differ from each other under the two transfection ratios. However, co-expression of an inactive P2X2 mutant and the wild type P2X3 subunit and vice versa resulted in characteristic distortions of the α,β-meATP concentration-response relationships, depending on which subunit was expressed in excess, suggesting that HEK293 cells express mixtures of (P2X2)1/(P2X3)2 and (P2X2)2/(P2X3)1 receptors. Whereas the allosteric modulators H+ and Zn2+ failed to discriminate between the two possible heterotrimeric receptor variants, the α,β-meATP-induced responses were blocked more potently by the competitive antagonist A317491, when the P2X2 subunit was expressed in deficit of the P2X3 subunit. Furthermore, blue-native PAGE analysis of P2X2 and P2X3 subunits co-expressed in Xenopus laevis oocytes and HEK293 cells revealed that plasma membrane-bound P2X2/3 receptors appeared in two clearly distinct heterotrimeric complexes: a (P2X2-GFP)2/(P2X3)1 complex and a (P2X2-GFP)1/(P2X3)2 complex. These data strongly indicate that the stoichiometry of the heteromeric P2X2/3 receptor is not fixed, but determined in a permutational manner by the relative availability of P2X2 and P2X3 subunits.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Neuropharmacology - Volume 99, December 2015, Pages 115-130
Journal: Neuropharmacology - Volume 99, December 2015, Pages 115-130
نویسندگان
Maria Kowalski, Ralf Hausmann, Julia Schmid, Anke Dopychai, Gabriele Stephan, Yong Tang, Günther Schmalzing, Peter Illes, Patrizia Rubini,