Maechler P, Wang H, Wollheim C B
Department of Internal Medicine, University Medical Centre, Geneva, Switzerland.
FEBS Lett. 1998 Feb 6;422(3):328-32. doi: 10.1016/s0014-5793(97)01618-9.
The second messenger role of ATP in insulin secretion was investigated in living INS-1 insulinoma cells. ATP-dependent luminescence was monitored in cells expressing high levels of firefly luciferase under the control of the tetracycline-dependent transactivator. The calibration of luminescence in permeabilized cells yielded similar ATP levels as those obtained in cell extracts with a conventional ATP assay. Stimulation of insulin secretion by glucose or methyl-succinate was correlated with rises of cellular ATP in simultaneous measurements. ATP generation was decreased by inhibition of the ADP-ATP translocase. This approach demonstrates the feasibility of defining the dynamic relationship between ATP and other parameters involved in metabolism-secretion coupling.
在活的INS-1胰岛素瘤细胞中研究了ATP在胰岛素分泌中的第二信使作用。在四环素依赖性反式激活因子控制下，对表达高水平萤火虫荧光素酶的细胞监测ATP依赖性发光。用传统的ATP测定法对透化细胞中的发光进行校准，得到的ATP水平与细胞提取物中获得的相似。在同步测量中，葡萄糖或甲基琥珀酸对胰岛素分泌的刺激与细胞ATP的升高相关。通过抑制ADP-ATP转位酶降低了ATP的生成。这种方法证明了定义ATP与代谢-分泌偶联中其他参数之间动态关系的可行性。
