Ferro A M, Oppenheimer N J
Proc Natl Acad Sci U S A. 1978 Feb;75(2):809-13. doi: 10.1073/pnas.75.2.809.
A preparation of poly(adenosine diphosphoribose) synthase obtained from pigeon liver nuclei has been used to make poly(adenosine diphosphoribose) with an average chain length of 20. Digestion of the purified poly(adenosine diphosphoribose) with snake venom phosphodiesterase (oligonucleate 5'-nucleotidohydrolase; EC 3.1.4.1) gave the monomer, 2'-(5"-phosphoribosyl)-5-AMP. After purification of the monomer on a Dowex-1 column, further digestion with alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum); EC 3.1.3.1] yielded the dephosphorylated product, 2'-ribosyl adenosine. Nuclear magnetic resonance spectra at 360 MHz of the 2'-ribosyl adenosine were obtained in [2H6]dimethylsulfoxide, which allows direct observation of the hydroxyl protons. These spectra show the absence of the adenosine 2'-hydroxyl proton, thus confirming the 2' position as the site of attachment of the ribose to the adenosine moiety. Comparison of the coupling constants and the chemical shifts of the ribose hydroxyl protons of 2'-ribosyl adenosine with the model compounds alpha- and beta-methylribofuranoside establishes an alpha (1" leads to 2') glycosidic linkage in the monomer. No evidence was found for heterogeneity in either the site of attachment or configuration of the linkage in the 2'-(5"-phosphoribosyl)-5'-AMP.
从鸽肝细胞核中获得的聚（腺苷二磷酸核糖）合成酶制剂已被用于制备平均链长为20的聚（腺苷二磷酸核糖）。用蛇毒磷酸二酯酶（寡核苷酸5'-核苷酸水解酶；EC 3.1.4.1）消化纯化的聚（腺苷二磷酸核糖），得到单体2'-（5''-磷酸核糖基）-5-AMP。在Dowex-1柱上纯化单体后，用碱性磷酸酶[正磷酸单酯磷酸水解酶（最适碱性）；EC 3.1.3.1]进一步消化，得到去磷酸化产物2'-核糖基腺苷。在[2H6]二甲基亚砜中获得了2'-核糖基腺苷在360 MHz的核磁共振谱，这使得可以直接观察羟基质子。这些谱显示腺苷2'-羟基质子不存在，从而证实2'位是核糖与腺苷部分连接的位点。将2'-核糖基腺苷的核糖羟基质子的耦合常数和化学位移与模型化合物α-和β-甲基呋喃核糖苷进行比较，确定单体中存在α（1''→2'）糖苷键。未发现2'-（5''-磷酸核糖基）-5'-AMP在连接位点或连接构型上存在异质性的证据。
