从大鼠组织中提取、纯化和鉴定3',5'-环磷酸胞苷
Extraction, purification and identification of cytidine 3',5'-cyclic monophosphate from rat tissues.
作者信息
Newton R P, Salih S G, Salvage B J, Kingston E E
出版信息
Biochem J. 1984 Aug 1;221(3):665-73. doi: 10.1042/bj2210665.
Abstract
The large-scale extraction and purification to homogeneity of cyclic CMP and its unequivocal identification are described. Rat liver, kidney, heart, spleen and lung tissues were subjected to a sequential purification procedure involving freeze-clamping, perchlorate extraction, alumina and boronate column chromatography, polyacrylamide-gel column electrophoresis and high-voltage paper electrophoresis. The purified sample co-chromatographed with authentic cyclic CMP on t.l.c. and high-pressure liquid chromatography and was positive in a cyclic CMP radio-immunoassay. The u.v., i.r. and p.m.r. spectra were each essentially identical with those of authentic cyclic CMP. Fast-atom bombardment of authentic cyclic CMP yielded a mass spectrum containing a molecular protonated ion: mass-ion-kinetic-energy scanning of this ion produced a spectrum unique to 3',5'-cyclic CMP. The extracted nucleotide produced an identical mass-ion-kinetic-energy spectrum.
摘要
本文描述了环状CMP的大规模提取、纯化至均一状态及其明确鉴定。对大鼠肝脏、肾脏、心脏、脾脏和肺组织进行了一系列纯化步骤，包括冷冻钳夹、高氯酸盐提取、氧化铝和硼酸盐柱色谱、聚丙烯酰胺凝胶柱电泳和高压纸电泳。纯化后的样品在薄层层析和高压液相色谱上与标准环状CMP共层析，并且在环状CMP放射免疫测定中呈阳性。紫外光谱、红外光谱和核磁共振光谱与标准环状CMP的光谱基本相同。对标准环状CMP进行快原子轰击产生的质谱包含一个分子质子化离子：对该离子进行质量-离子-动能扫描产生了3',5'-环状CMP特有的光谱。提取的核苷酸产生了相同的质量-离子-动能光谱。
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