Kraft A S, Appling C, Berkow R L
Biochem Biophys Res Commun. 1987 Apr 14;144(1):393-401. doi: 10.1016/s0006-291x(87)80523-5.
In this report, we demonstrate that HL-60 nuclei isolated in calcium but not EGTA containing buffers specifically bind PE and express approximately 37,000 receptor sites/nucleus. Nuclear phorbol ester binding is lost by isolation in the absence of calcium, but can be repleted by the addition of partially purified protein kinase C and calcium. When HL-60 cells are treated with bryostatin 1, a compound which activates protein kinase C in a similar fashion to phorbol esters but does not induce differentiation of HL-60 cells, and nuclei are isolated in the presence of EGTA, these nuclei continue to bind phorbol esters. These experiments suggest that HL-60 nuclei bind PE in vitro, and that compounds that activate protein kinase C may increase nuclear binding of PE in situ.
在本报告中,我们证明,在含有钙而非乙二醇双乙醚四乙酸(EGTA)的缓冲液中分离得到的HL-60细胞核可特异性结合佛波酯(PE),且每个细胞核表达约37,000个受体位点。在无钙条件下分离时,细胞核对佛波酯的结合能力丧失,但添加部分纯化的蛋白激酶C和钙后可恢复。当用苔藓抑素1处理HL-60细胞(一种以与佛波酯类似的方式激活蛋白激酶C但不诱导HL-60细胞分化的化合物),并在EGTA存在的情况下分离细胞核时,这些细胞核仍继续结合佛波酯。这些实验表明,HL-60细胞核在体外可结合PE,且激活蛋白激酶C的化合物可能会增加原位PE的核结合。