Culcasi Marcel, Thétiot-Laurent Sophie, Atteia Ariane, Pietri Sylvia
UMR 7273, CNRS, Equipe Sondes Moléculaires en Biologie et Stress Oxydant, Centre scientifique de Saint-Jérôme, Institut de Chimie Radicalaire, Aix-Marseille Université, Service 522, Avenue Escadrille Normandie-Niemen, Marseille Cedex 20, 13397, France.
Methods Mol Biol. 2015;1265:135-47. doi: 10.1007/978-1-4939-2288-8_11.
(31)P nuclear magnetic resonance (NMR) is a unique technique to monitor noninvasively the energetics of living systems at real time through the detection of a variety of phosphorylated metabolites. Using adequately designed α-aminophosphonates as external probes, we have shown earlier that (31)P NMR can also give access simultaneously to the accurate pH of cytosolic and acidic compartments in normal and stressed cultured cells or isolated perfused organs, a feature that was not possible using endogenous inorganic phosphate as the probe. More recently, we obtained a series of derivatives of these new pH probes that incorporate a triphenylphosphonium cation as a specific vector to the mitochondrion. Here, we describe the synthesis, (31)P NMR pH titrating properties in buffers, and application in cultures of the green alga Chlamydomonas reinhardtii of two of these mitochondria-targeted pH probes in comparison with one nonvectorized, yet still informative α-aminophosphonate.
磷-31核磁共振（NMR）是一种独特的技术，可通过检测多种磷酸化代谢物实时无创监测生物系统的能量学。我们早前已表明，使用设计得当的α-氨基膦酸盐作为外部探针，磷-31 NMR还能够同时获取正常和应激培养细胞或离体灌注器官中胞质和酸性区室的准确pH值，而使用内源性无机磷酸盐作为探针则无法做到这一点。最近，我们获得了这些新型pH探针的一系列衍生物，它们含有一个三苯基鏻阳离子作为靶向线粒体的特定载体。在此，我们描述了其中两种靶向线粒体的pH探针的合成、在缓冲液中的磷-31 NMR pH滴定特性以及在莱茵衣藻培养中的应用，并与一种无载体但仍具信息价值的α-氨基膦酸盐作比较。
