Kozarich J W, Chinault A C, Hecht S M
Biochemistry. 1975 Mar 11;14(5):981-8. doi: 10.1021/bi00676a017.
A procedure has been outlined for the synthesis of ribonucleoside 3'-di- and -triphosphates. The synthetic scheme involves the conversion of a ribonucleoside 3'-monophosphate to its 2'-(5'-di)-O-(1-methoxyethyl) derivative, followed by successive treatments of the blocked ribonucleotide with 1,1'-carbonyldiimidazole and mono(tri-n-butylammonium) phosphate or pyrophosphate. The resulting ribonucleoside 3'-di- and -triphosphate derivatives are then deblocked by treatment with dilute aqueous acetic acid, pH 3.0. The use of this procedure is illustrated for adenosine 3'-monophosphate, which has been converted to its corresponding 3'-di- and -triphosphates in 61% overall yield. The decomposition of adenosine 3'-di- and -triphosphates to adenosine 2'-monophosphate, adenosine 3'-monophosphate, and adenosine cyclic 2',3'-monophosphate as a function of pH at 100 degrees has been studied as has the attempted polymerization of adenosine 3'-diphosphate with polynucleotide phosphorylase. Also prepared was guanosine 5'-diphosphate 3'-diphosphate (guanosine tetraphosphate; ppGpp), which was accessible via treatment of 2'-O-(1-methoxyethyl)guanosine 5'-monophosphate 3'-monophosphate with the phosphorimidazolidate of mono(tri-n-butyl ammonium) phosphate. The resulting blocked tetraphosphate was deblocked in dilute aqueous acetic acid to afford ppGpp in an overall yield of 18%.
已概述了一种合成核糖核苷3'-二磷酸和三磷酸的方法。合成方案包括将核糖核苷3'-单磷酸转化为其2'-(5'-二)-O-(1-甲氧基乙基)衍生物，然后用1,1'-羰基二咪唑和磷酸单（三正丁基铵）或焦磷酸对封闭的核糖核苷酸进行连续处理。然后用pH 3.0的稀醋酸水溶液处理，使所得的核糖核苷3'-二磷酸和三磷酸衍生物脱保护。以腺苷3'-单磷酸为例说明了该方法的应用，其已转化为相应的3'-二磷酸和三磷酸，总产率为61%。研究了腺苷3'-二磷酸和三磷酸在100℃下分解为腺苷2'-单磷酸、腺苷3'-单磷酸和腺苷环2',3'-单磷酸与pH的关系，以及腺苷3'-二磷酸与多核苷酸磷酸化酶的聚合尝试。还制备了鸟苷5'-二磷酸3'-二磷酸（鸟苷四磷酸；ppGpp），可通过用磷酸单（三正丁基铵）的磷酰咪唑盐处理2'-O-(1-甲氧基乙基)鸟苷5'-单磷酸3'-单磷酸来获得。所得的封闭四磷酸在稀醋酸水溶液中脱保护，得到ppGpp，总产率为18%。
