Fluorescent kabiramides: New probes to quantify actin in vitro and in vivo
Fluorescent kabiramides: New probes to quantify actin in vitro and in vivo
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DOI：
10.1021/bc050006j
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发表时间：
2005-11-01
影响因子：
4.7
通讯作者：
Marriott, G
中科院分区：
文献类型：
--
作者：
Petchprayoon, C;Suwanborirux, K;Marriott, G
我们目前的设计，合成，和生化和光谱特性的五个功能性的卡比胺C（KabC），凝溶胶蛋白的小分子仿生荧光共轭物。KabC的四甲基罗丹明（TMR）、罗丹酚绿色（RG）、IC 5、达泊昔（DAP）和荧光素二酯（FDE）缀合物以1：1的复合物在倒刺末端特异性结合肌动蛋白。这些探针显示以与未修饰的KabC不可区分的方式起作用。这些KabC探针的荧光发射的各种模式，包括荧光各向异性和荧光共振能量转移，用于开发用于快速测定溶液中G-肌动蛋白浓度的测定。TMR-KabC和FDE-KabC探针是细胞可渗透的，并提供关于活细胞内肌动蛋白丝的分布和动力学的独特成像信息。
We present the design, synthesis, and biochemical and spectroscopic characterization of five functional fluorescent conjugates of kabiramide C (KabC), a small molecule biomimetic of gelsolin. The tetramethylrhodamine (TMR), rhodol green (RG), IC5, dapoxyl (DAP), and fluorescein diester (FDE) conjugates of KabC bind specifically to actin at the barbed end in a 1:1 complex. These probes are shown to function in an indistinguishable manner to the unmodified KabC. Various modalities of the fluorescence emission of these KabC probes, including fluorescence anisotropy and fluorescence resonance energy transfer, are used for the development of assays for the rapid determination of G-actin concentration in solution. The TMR-KabC and FDE-KabC probes are cell permeable and provide unique imaging information on the distribution and dynamics of actin filament within living cells.