从组织样本中的胸腺嘧啶核苷、电解质和多价核苷酸中分离环磷酸腺苷和环磷酸鸟苷。

Separation of cyclic AMP and cyclic GMP from thymidine, electrolytes and polyvalent nucleotides in tissue samples.

作者信息

Roberson G M, Barnes L D

出版信息

Biochim Biophys Acta. 1978 Nov 15;544(1):20-8. doi: 10.1016/0304-4165(78)90205-2.

Abstract

Cyclic AMP and cyclic GMP can be separated from thymidine and its possible metabolites, electrolytes, and polyvalent nucleotides using columns of acidic alumina. Electrolytes and thymidine are not adsorbed on acidic alumina at pH 4.4 while cyclic nucleotides and polyvalent nucleotides are adsorbed at this pH. Cyclic AMP and cyclic GMP are eluted together from acidic alumina with 0.2 M ammonium formate (pH 6.0) and the polyvalent nucleotides remain adsorbed. The cyclic nucleotides are separated by chromatography on Dowex AG 1 X 8 resin. Recovery is 60--64% for cyclic AMP and cyclic GMP isolated from renal tissue samples. This methodology permits the separation of tritiated thymidine from cyclic nucleotides which are present in tissue preparations used in studies on the role of cyclic nucleotides in cellular growth.

摘要

使用酸性氧化铝柱，可将环磷酸腺苷（cAMP）和环磷酸鸟苷（cGMP）与胸腺嘧啶核苷及其可能的代谢产物、电解质和多价核苷酸分离。在pH 4.4时，电解质和胸腺嘧啶核苷不会吸附在酸性氧化铝上，而环核苷酸和多价核苷酸在此pH值下会被吸附。cAMP和cGMP用0.2M甲酸铵（pH 6.0）从酸性氧化铝上一起洗脱下来，多价核苷酸仍被吸附。通过在Dowex AG 1 X 8树脂上进行色谱分离可将环核苷酸分离。从肾组织样品中分离得到的cAMP和cGMP的回收率为60% - 64%。这种方法能够将氚标记的胸腺嘧啶核苷与环核苷酸分离，环核苷酸存在于用于研究环核苷酸在细胞生长中作用的组织制剂中。