Gruenwedel D W, Heskett M G, Lammert J E
Biochim Biophys Acta. 1975 Aug 6;402(1):7-19. doi: 10.1016/0005-2787(75)90364-0.
Dextran gels of varying porosites (Sephadex G series) were chemically modified so as to contain covalently bound monofunctional mercury. Mercurated Sephadex of the porosity G-25 was then used to fractionate mixtures of mono- and dinucleotides into the constituent components. Separation is based on the affinity of the nitrogen binding sites of the purine and pyrimidine derivatives for organomercurial Hg+. Thus, a mixture composed of the four mononucleotides Cyd-5'-P, Ado-3'-P, Guo-2'(3')-P, dThd-5'-P and of the four dinucleotides Cyd-P-Cyd, Ado-P-Ado, Guo-P-Urd, and Urd-P-Urd could be separated into eight well-resolved fractions by using a combination Tris-bicarbonate/carbonate buffer system of increasing pH as an eluent. Complete separation was also achieved when a mixture of Ado 3:5'-P, Ado 5'-P, Ado-5'-PP, and Ado-5'-PPP was fractionated on mercurated Sephadex G-25. Again, Tris-bicarbonate/carbonate buffer served as an eluent. Lastly, fractionation can also be performed at a constant pH by offering other suitable ligands, for instance Cl-, that will compete with nucleotides for monofunctional Hg+. The fractionation behavior of mercurated Sephadex G-25 can be fully understood on the basis of the complexing properties of monofunctional Hg+. This has been shown by calculating the net retention volume ratios of several nucleotides with the help of the known interaction parameters of corresponding nucleosides with CH3 HgOH and by comparing the predicted ratios with the experimentally measured ones. Finally, the acid-base properties of mercurated Sephadex G-25 as well as its affinity for chloride and iodide ions have been determined. The data agree quite well with those known for CH3 HgOH.
对不同孔隙率的葡聚糖凝胶（葡聚糖凝胶G系列）进行化学修饰，使其含有共价结合的单功能汞。然后用孔隙率为G - 25的汞化葡聚糖凝胶将单核苷酸和二核苷酸混合物分离成其组成成分。分离基于嘌呤和嘧啶衍生物的氮结合位点对有机汞Hg⁺的亲和力。因此，由四种单核苷酸胞苷 - 5'-磷酸（Cyd - 5'-P）、腺苷 - 3'-磷酸（Ado - 3'-P）、鸟苷 - 2'(3') - 磷酸（Guo - 2'(3') - P）、脱氧胸苷 - 5'-磷酸（dThd - 5'-P）以及四种二核苷酸胞苷 - 磷酸 - 胞苷（Cyd - P - Cyd）、腺苷 - 磷酸 - 腺苷（Ado - P - Ado）、鸟苷 - 磷酸 - 尿苷（Guo - P - Urd）和尿苷 - 磷酸 - 尿苷（Urd - P - Urd）组成的混合物，通过使用pH值递增的Tris - 碳酸氢盐/碳酸盐缓冲系统作为洗脱剂，可分离成八个分离良好的组分。当腺苷3:5'-磷酸（Ado 3:5'-P）、腺苷5'-磷酸（Ado 5'-P）、腺苷 - 5'-二磷酸（Ado - 5'-PP）和腺苷 - 5'-三磷酸（Ado - 5'-PPP）的混合物在汞化葡聚糖凝胶G - 25上进行分离时，也实现了完全分离。同样，Tris - 碳酸氢盐/碳酸盐缓冲液用作洗脱剂。最后，通过提供其他合适的配体，例如Cl⁻，它会与核苷酸竞争单功能Hg⁺，也可以在恒定pH下进行分离。基于单功能Hg⁺的络合性质，可以充分理解汞化葡聚糖凝胶G - 25的分离行为。通过借助相应核苷与CH₃HgOH的已知相互作用参数计算几种核苷酸的净保留体积比，并将预测值与实验测量值进行比较，已证明了这一点。最后，测定了汞化葡聚糖凝胶G - 25的酸碱性质及其对氯离子和碘离子的亲和力。数据与CH₃HgOH的已知数据相当吻合。
