Amine-reactive forms of a luminescent diethylenetriaminepentaacetic acid chelate of terbium and europium: Attachment to DNA and energy transfer measurements
Amine-reactive forms of a luminescent diethylenetriaminepentaacetic acid chelate of terbium and europium: Attachment to DNA and energy transfer measurements
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DOI：
10.1021/bc960085m
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发表时间：
1997-03-01
影响因子：
4.7
通讯作者：
Selvin, PR
中科院分区：
文献类型：
--
作者：
Li, M;Selvin, PR
合成了一种异硫氰酸酯形式的稀土络合物，它在与Tb或Eu结合时具有很高的发光性能。该络合物由二乙三胺五乙酸共价连接到生色团7-氨基-4-甲基-2(1H)-喹啉酮(CS124)和L-对氨基苯丙氨酸，其中芳香胺进一步转化为异硫氰酸酯基团。乙二胺也被用来代替氨基苯丙氨酸，但由脂肪胺形成的异硫氰酸酯的活性明显较低。已经在甘氨酸和胺修饰的DNA寡聚体的5‘端进行了定位连接。此外，作为一种与大分子偶联的替代方法，DTPA酸酐-CS124可用于在去碱基保护的合成DNA低聚物上与5‘胺基发生特异性反应。在这两种情况下，合成和纯化都相对简单，发光性质有利于几种应用，包括作为非同位素标记，作为成像和诊断中荧光团的长寿命替代品，特别是作为发光共振能量转移的供体。能量转移测量结果与以前报道的使用不同附着机制的测量结果一致。
An isothiocyanate form of a lanthanide chelate which is highly luminescent when bound to terbium or europium has been synthesized. The chelate consists of diethylenetriaminepentaacetic acid (DTPA) covalently joined to a chromophore, 7-amino-4-methyl-2(1H)-quinolinone (cs124), and to L-p-aminophenylalanine, in which the aromatic amine was further converted to an isothiocyanate group. Ethylenediamine was also used in place of aminophenylalanine, but the isothiocyanate formed from the aliphatic amine was significantly less reactive. Site-specific attachments to triglycine and to the 5' ends of amine-modified DNA oligomers have been made. In addition, as an alternative method of coupling to macromolecules, DTPA anhydride-cs124 can be used to react specifically with a 5' amine group on base-deprotected synthetic DNA oligomers. Synthesis and purification is relatively straightforward in both cases, and luminescent properties are favorable for several applications, including as nonisotopic labels, as long-lifetime alternatives to fluorophores in imaging and diagnostics and particularly as donors in luminescence resonance energy transfer. Energy transfer measurements are consistent with previously reported measurements using different attachment mechanisms.