کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1948929 | 1054718 | 2006 | 7 صفحه PDF | دانلود رایگان |
LIBRA is a fluorescent biosensor of inositol 1,4,5-trisphosphate (IP3) and is composed of the ligand-binding domain of the rat type 3 IP3 receptor and cyan and yellow fluorescent proteins. We examined the responses of LIBRA and its IP3-insensitive mutant LIBRA-N to compounds known to inhibit IP3-induced Ca2+ release. Heparin, a competitive antagonist of IP3 receptors, increased the emission ratio of LIBRA but not that of LIBRA-N. In contrast, 2-aminoethoxydiphenyl borate, a known non-competitive inhibitor of IP3 receptor, decreased the emission ratios of both LIBRA and LIBRA-N. Thus, the concurrent use of LIBRA-N with LIBRA identifies nonspecific responses. These results indicate that LIBRA and its mutant control can be used to detect specific agonists and antagonists of IP3 receptors. We also demonstrate the utility of LIBRA and LIBRA-N in discriminating between specific and nonspecific responses in intact cells.
Journal: Biochimica et Biophysica Acta (BBA) - General Subjects - Volume 1760, Issue 8, August 2006, Pages 1274–1280