公开(公告)号：US09651494B2

公开(公告)日：2017-05-16

申请号：US14118836

申请日：2012-05-21

申请人： Evan Walker Miller ,  Roger Y. Tsien

发明人： Evan Walker Miller ,  Roger Y. Tsien

IPC分类号： G01N21/64 ,  G01N33/50 ,  G01N33/542 ,  C09K11/06

CPC分类号： G01N21/6486 ,  C09K11/06 ,  C09K2211/1007 ,  C09K2211/1011 ,  C09K2211/1088 ,  G01N33/5008 ,  G01N33/542 ,  G01N2500/10

摘要： Compounds and methods for determining transmembrane potential, monitoring changes in transmembrane potential, and/or drug screening are provided. In one aspect, compounds of the invention have a structure according to the formula: E-M-A, wherein A is a fluorophore, selected from xanthenes, coumarins, cyanines, bimanes, and difluoroboradizaindacenes, charged at physiological pH; M is a molecular wire; and E is a hydrophobic moiety, wherein A and E are capable of being involved in a photo-induced, intramolecular electron transfer that quenches the fluorescence of A in response to a voltage condition. When in use, compounds of the invention are membrane-impermeant and oriented within the cell membrane such that the charged moiety localizes at the outer leaflet of the lipid bilayer and the hydrophobic moiety and molecular wire associate with the hydrophobic portion of the lipid bilayer. The rate of electron transfer, fluorescence intensity, and quenching are altered in response to changes in transmembrane potential.

公开(公告)号：US20140093907A1

公开(公告)日：2014-04-03

申请号：US14118836

申请日：2012-05-21

申请人： Evan Walker Miller ,  Roger Y. Tsien

发明人： Evan Walker Miller ,  Roger Y. Tsien

IPC分类号： G01N21/64

CPC分类号： G01N21/6486 ,  C09K11/06 ,  C09K2211/1007 ,  C09K2211/1011 ,  C09K2211/1088 ,  G01N33/5008 ,  G01N33/542 ,  G01N2500/10

摘要： Compounds and methods for determining transmembrane potential, monitoring changes in transmembrane potential, and/or drug screening are provided. In one aspect, compounds of the invention have a structure according to the formula: E-M-A, wherein A is a fluorophore, selected from xanthenes, eoumarins, cyanines, bimanes, and difluoroboradizaindacenes, charged at physiological pH; M is a molecular wire; and E is a hydrophobic moiety, wherein A and E are capable of being involved in a photo-induced, intramolecular electron transfer that quenches the fluorescence of A in response to a voltage condition. When in use, compounds of the invention are membrane-impermeant and oriented within the cell membrane such that the charged moiety localizes at the outer leaflet of the lipid bilayer and the hydrophobic moiety and molecular wire associate with the hydrophobic portion of the lipid bilayer. The rate of electron transfer, fluorescence intensity, and quenching are altered in response to changes in transmembrane potential.

摘要翻译： 提供了确定跨膜电位的化合物和方法，监测跨膜电位的变化和/或药物筛选。 在一个方面，本发明的化合物具有下式的结构：其中A是荧光团，选自在生理pH下加入的呫吨，香豆素，花青，双马来酰亚胺和二氟硼烷二恶烷; M是分子线; 并且E是疏水部分，其中A和E能够参与光诱导的分子内电子转移，其响应于电压条件淬灭A的荧光。 当使用时，本发明的化合物是膜不渗透的并且在细胞膜内定向，使得带电部分定位在脂质双层的外部小叶上，并且疏水部分和分子线与脂质双层的疏水部分缔合。 响应于跨膜电位的变化，电子转移速率，荧光强度和淬灭速率发生变化。