Auchus R J, Kaiser S L, Majerus P W
Proc Natl Acad Sci U S A. 1987 Mar;84(5):1206-9. doi: 10.1073/pnas.84.5.1206.
We have developed a method for synthesis of inositol 1,2-(cyclic)-4,5-trisphosphate from inositol 1,4,5-trisphosphate using a water-soluble carbodiimide. We obtained 1-1.5 mumol of the inositol cyclic trisphosphate starting with 5 mumol of inositol 1,4,5-trisphosphate. The cyclized product was isolated by HPLC on Partisil SAX. The identity of the cyclic product was verified by its hydrolysis to inositol 1,4,5-trisphosphate in acid and by its conversion to 1,2-(cyclic)-4-bisphosphate by a specific 5-phosphomonoesterase from platelets. We also identified the product by 31P NMR spectroscopy, which showed a peak at 17.2 ppm, characteristic of a five-membered cyclic phosphodiester ring, and peaks at 4.1 ppm and 0.8 ppm, indicative of phosphomonoesters. This relatively simple method for producing inositol 1,2-(cyclic)-4,5-trisphosphate will facilitate studies of the physiology of this compound in signal transduction.
我们开发了一种使用水溶性碳二亚胺从肌醇1,4,5-三磷酸合成肌醇1,2-(环)-4,5-三磷酸的方法。以5 μmol肌醇1,4,5-三磷酸为起始原料，我们获得了1 - 1.5 μmol的肌醇环三磷酸。环化产物通过在Partisil SAX上进行高效液相色谱法分离。通过在酸性条件下水解为肌醇1,4,5-三磷酸以及通过血小板特异性5-磷酸单酯酶将其转化为1,2-(环)-4-二磷酸，验证了环化产物的身份。我们还通过31P核磁共振光谱法鉴定了该产物，其在17.2 ppm处显示一个峰，这是五元环磷酸二酯环的特征峰，在4.1 ppm和0.8 ppm处有峰，表明是磷酸单酯。这种相对简单的生产肌醇1,2-(环)-4,5-三磷酸的方法将有助于研究该化合物在信号转导中的生理学特性。
