synthesis and in vitro evaluation of dioxopyrrolopyrroles as potential low-affinity fluorescent indicators合成和体外评价dioxopyrrolopyrroles作为潜在的低亲和力荧光指标.pdfVIP

Vol. 06 INTERNATIONAL JOURNAL OF PHOTOENERGY 2004 Synthesis and in vitro evaluation of dioxopyrrolopyrroles as potential low-affinity fluorescent Ca2+ indicators Nesibe Avcıba¸sı,1,2 Mario Smet,1 Bert Metten,1 Wim Dehaen,1,† Frans C. De Schryver,1 Geert Bultynck,3 Geert Callewaert,3 Humbert De Smedt,3 Ludwig Missiaen,3 and Noël Boens1,‡ 1 Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001 Heverlee, Belgium 2 Department of Chemistry, Faculty of Science, Ege University, 35100 Bornova, Izmir, Turkey 3 Laboratory of Physiology, Katholieke Universiteit Leuven, Campus Gasthuisberg O N, Herestraat 49, 3001 Heverlee, Belgium Abstract. Three new low-affinity fluorescent Ca2+ indicators excitable with visible light, namely 3-phenyl-6-(4-(3-carboxymethoxy-4-(N,N- dicarboxymethylamino)phenyl)phenyl)-2,5-dicarboxymethyl-1,4- dihydropyrrolo[3,4-c]pyrrole-1,4-dione (DPP1), 3-phenyl-6-(5-(3-carboxymethoxy-4-(N,N-dicarboxymethyl- amino)phenyl)thien-2-yl)-2,5-dicarboxymethyl-1,4-dihydropyrrolo[3,4-c]pyrrole-1,4-dione (DPP2) and 3- (thien-2-yl)-6-(5-(3-carboxymethoxy-4- (N,N-dicarboxymethylamino)phenyl)thien-2-yl)-2,5-dicarboxymethyl- 1,4-dihydropyrrolo[3,4-c]pyrrole-1,4-dione (DPP3) have been synthesized and evaluated for their Ca2+ binding properties via fluorimetric titrations. The in vitro dissociation constant Kd measured at 21 ◦C in 100 mM KCl buffered solution, pH 7.05, for the Ca2+–DPP1 complex is 10 µM; for Ca2+–DPP2 and Ca2+–DPP3 a Kd value of 20 µM is fou