Synthesis and characterization of conjugates formed between periodate-oxidized ribonucleotides and amine-containing fluorophores.
Synthesis and characterization of conjugates formed between periodate-oxidized ribonucleotides and amine-containing fluorophores.
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高碘酸盐氧化核糖核苷酸和含胺荧光团之间形成的缀合物的合成和表征。
DOI：
10.1021/bc00029a010
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发表时间：
1994
影响因子：
4.7
通讯作者：
Parkhurst,LJ
中科院分区：
文献类型：
--
作者：
Hileman,RE;Parkhurst,KM;Gupta,NK;Parkhurst,LJ
介绍了新的荧光标记的GDP和ATP衍生物的合成和纯化。荧光基团最初通过含胺的连接臂偶联到高碘酸盐氧化的核苷酸。初始产物的还原主要产生一个六元的似morpholine环。荧光素标记的GDP、罗丹明标记的GDP和荧光素标记的ATP通过吸光度光谱和薄层色谱进行表征。对一个单核苷酸衍生物进行NMR和FAB-MS研究，该衍生物是由高尿酸盐氧化的鸟苷和苄胺反应形成的，随后进行还原，以确定对核糖部分的修饰。鸟苷-苄胺缀合物的合成导致了分离产物的混合物。主要产物（70%）将核糖部分转化为没有羟基的六元morpholine样环，次要产物（30%）导致具有一个羟基的开环结构。当只使用NaCNBHg作为还原剂时，只生成具有仿吗啡环的产物。这些探针准备用于真核起始因子-2与哺乳动物蛋白质合成起始的其他组分相互作用的溶液研究。
The synthesis and purification of new fluorescently labeled derivatives of GDP and ATP are described. The fluorescent groups are coupled initially through amine-containing linker arms to periodate-oxidized nucleotides. Reduction of the initial product yields primarily a six-membered morpholine-like ring. Fluorescein-labeled GDP, rhodamine-labeled GDP, and fluorescein-labeled ATP were characterized by absorbance spectroscopy and TLC. NMR and FAB-MS studies were carried out on a single nucleotide derivative formed by reacting periodate-oxidized guanosine and benzylamine with subsequent reduction to establish the modification to the ribose moiety. The synthesis of the guanosine—benzylamine conjugate led to a mixture of products that were separated. The predominant product (70%) resulted in conversion of the ribose moiety to a six-membered morpholine-like ring having no hydroxyl group, and the minor product (30%) resulted in an open ring structure having one hydroxyl. When NaCNBHg was used as the sole reductant, only the product with the morpholinelike ring was formed. These probes were prepared for use in solution studies of the interactions of eukaryotic initiation factor-2 with other components of mammalian protein synthesis initiation.