Identification of two oligodeoxyribonucleotide binding proteins on plasma membranes of human cell lines.
Identification of two oligodeoxyribonucleotide binding proteins on plasma membranes of human cell lines.
复制标题
人类细胞系质膜上两种寡脱氧核糖核苷酸结合蛋白的鉴定。
DOI：
10.1016/0006-2952(95)02198-1
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发表时间：
1996
影响因子：
5.8
通讯作者：
Cheng,YC
中科院分区：
文献类型：
--
作者：
Yao,GQ;Corrias,S;Cheng,YC
用光标记技术在人HL-60、HepG 2、H1和KB细胞质膜上发现了两个分子量约为100-110 kDa的寡脱氧核苷酸（oligodN）结合蛋白。用非离子去污剂溶解细胞膜并不干扰这两种蛋白质与oligodNs的结合，并且这两种蛋白质都容易受到丝氨酸蛋白酶作用的影响。Scatchard图分析显示磷酸二酯（PO）21-mer寡脱氧胞苷的Kd为60 nM，HepG 2细胞的结合位点数约为1.2 × 106个/细胞。硫代磷酸酯（PS）和PO寡核苷酸都能与这两种蛋白结合，PS寡核苷酸的结合亲和力远强于PO寡核苷酸。与oligodNs的结合受到反应的离子强度的影响。硫酸葡聚糖、tRNA和双链DNA抑制寡脱氧核糖核酸的结合，而ATP、ADP、AMP和TTP没有影响。鉴于它们对寡脱氧核苷酸的高亲和力，这些膜蛋白可能在寡脱氧核苷酸的作用中发挥重要作用。
Two oligodeoxyribonucleotide (oligodN) binding proteins of approximately 100–110 kDa were identified in the plasma membranes of human HL-60, HepG2, H1, and KB cells by a photolabeling technique. Solubilization of cellular membranes with a nonionic detergent did not interfere with the binding of these two proteins to oligodNs, and both proteins were susceptible to serine protease action. The binding affinities of these two proteins to oligodNs were found to be similar; Scatchard plot analysis revealed the Kdfor phosphodiester (PO) 21-mer oligodeoxycytidine to be 60 nM and binding sites numbered approximately 1.2 × 106/cell for HepG2 cells. Both phosphorothioate (PS) and PO oligodNs could bind to these two proteins with the binding affinity for PS oligodNs being much stronger than that for PO oligodNs. The binding to oligodNs was affected by the ionic strength of the reaction. Dextran sulfate, tRNA, and double-stranded DNA inhibited the binding of oligodNs, whereas ATP, ADP, AMP, and TTP had no effect. Given their high affinity for oligodNs, these membrane proteins may play an important role in the action of oligodNs.