一种通过亲和洗脱色谱法检测完整细胞中腺苷酸环化酶的简单测定方法。

A simple assay for adenylate cyclase in intact cells by affinity elution chromatography.

作者信息

Sinha A K, Colman R W

出版信息

Biochem J. 1978 Sep 15;174(3):699-702. doi: 10.1042/bj1740699.

DOI:10.1042/bj1740699

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1185972/

Abstract

A method using the principle of affinity elution chromatography is described for the assay of adenylate cyclase in intact human platelets. By incubating platelet-rich plasma in the presence of radioactively labelled adenine, the ATP pool of the cells was prelabelled. Formation of labelled cyclic AMP from ATP was determined by extracting the platelets with HC1O4. After removal of the latter as KC1O4, the extract containing cyclic AMP and other adenine nucleotides was adsorbed in a NN-diethyl-N-2-hydroxypropylamino (QAE)-cellulose column. The column was washed, and subsequently cyclic AMP was specifically eluted with a cyclic AMP-dependent protein kinase and the radioactivity of the eluate was determined.

摘要

本文描述了一种利用亲和洗脱色谱原理来测定完整人血小板中腺苷酸环化酶的方法。通过在放射性标记的腺嘌呤存在下孵育富含血小板的血浆，细胞的ATP池被预先标记。用高氯酸提取血小板来测定由ATP形成的标记环磷酸腺苷。在以高氯酸钾形式除去高氯酸后，含有环磷酸腺苷和其他腺嘌呤核苷酸的提取物被吸附到NN-二乙基-N-2-羟丙基氨基（QAE）-纤维素柱上。对该柱进行洗涤，随后用一种环磷酸腺苷依赖性蛋白激酶特异性洗脱环磷酸腺苷，并测定洗脱液的放射性。

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Adenyl cyclase. II. The enzymatically catalyzed formation of adenosine 3',5'-phosphate and inorganic pyrophosphate from adenosine triphosphate.腺苷酸环化酶。II. 由三磷酸腺苷酶催化形成3',5'-磷酸腺苷和无机焦磷酸。

J Biol Chem. 1962 Apr;237:1228-32.

2

THE ACTION OF EPINEPHRINE AND THE ROLE OF THE ADENYL CYCLASE SYSTEM IN HORMONE ACTION.肾上腺素的作用及腺苷酸环化酶系统在激素作用中的角色。

Recent Prog Horm Res. 1965;21:623-46.

3

Cyclic 3',5'-AMP and the lipolytic effects of hormones on adipose tissue.

Pharmacol Rev. 1966 Mar;18(1):237-41.

4

The separation and evaluation of puric bases and their nucleosides and nucleotides by thin-layer chromatography.通过薄层色谱法分离和评估嘌呤碱及其核苷和核苷酸。

Clin Chim Acta. 1974 May 31;53(1):69-77. doi: 10.1016/0009-8981(74)90353-2.

5

A simple and sensitive saturation assay method for the measurement of adenosine 3':5'-cyclic monophosphate.一种用于测量3':5'-环磷酸腺苷的简单且灵敏的饱和测定方法。

Biochem J. 1971 Feb;121(3):561-2. doi: 10.1042/bj1210561.

6

The influence on platelet aggregation of drugs that affect the accumulation of adenosine 3':5'-cyclic monophosphate in platelets.影响血小板中3':5'-环磷酸腺苷积累的药物对血小板聚集的影响。

Biochem J. 1971 Jan;121(2):185-96. doi: 10.1042/bj1210185.

7

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Biochemistry. 1970 Oct 13;9(21):4223-9. doi: 10.1021/bi00823a026.

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