来自纯化人T淋巴细胞的环核苷酸磷酸二酯酶的高压液相色谱法

High pressure liquid chromatography of cyclic nucleotide phosphodiesterase from purified human T-lymphocytes.

作者信息

Robicsek S A, Krzanowski J J, Szentivanyi A, Polson J B

机构信息

Department of Pharmacology and Therapeutics, University of South Florida, College of Medicine, Tampa 33612-4799.

出版信息

Biochem Biophys Res Commun. 1989 Aug 30;163(1):554-60. doi: 10.1016/0006-291x(89)92173-6.

DOI:10.1016/0006-291x(89)92173-6

PMID:2549997

Abstract

The cyclic nucleotide phosphodiesterase (EC 3.1.4.17) in extracts of purified human peripheral blood T-lymphocytes was examined by ion exchange high pressure liquid chromatography. Four peaks of activity were isolated. The first peak of activity selectively hydrolyzed cyclic GMP. The following 3 peaks of activity (Ia, IIa and IIIa) were selective for cyclic AMP. The selective low Km cyclic AMP-phosphodiesterase inhibitor, Ro 20-1724 (d,1-1,4-[3-butoxy-4-methoxybenzyl]-2-imidazolidinone), did not inhibit the activity in Ia whereas it did inhibit the activity in IIa and IIIa (IC50 = 17 microM). The authors conclude that ion exchange high pressure liquid chromatography described in this communication is a useful method for the isolation of different forms of cyclic nucleotide phosphodiesterase activity from human T-lymphocytes.

摘要

通过离子交换高压液相色谱法检测了纯化的人外周血T淋巴细胞提取物中的环核苷酸磷酸二酯酶（EC 3.1.4.17）。分离出了四个活性峰。第一个活性峰选择性地水解环鸟苷酸。接下来的三个活性峰（Ia、IIa和IIIa）对环腺苷酸具有选择性。选择性低Km环腺苷酸磷酸二酯酶抑制剂Ro 20-1724（d，1-1，4-[3-丁氧基-4-甲氧基苄基]-2-咪唑啉酮）不抑制Ia中的活性，而抑制IIa和IIIa中的活性（IC50 = 17 microM）。作者得出结论，本通讯中描述的离子交换高压液相色谱法是从人T淋巴细胞中分离不同形式环核苷酸磷酸二酯酶活性的有用方法。