Increased expression of the inositol 1,4,5-trisphosphate receptor in human leukaemic (HL-60) cells differentiated with retinoic acid or dimethyl sulphoxide.
Increased expression of the inositol 1,4,5-trisphosphate receptor in human leukaemic (HL-60) cells differentiated with retinoic acid or dimethyl sulphoxide.
复制标题
用视黄酸或二甲亚砜分化的人白血病 (HL-60) 细胞中肌醇 1,4,5-三磷酸受体的表达增加。
DOI：
10.1042/bj2800205
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Autieri,M
中科院分区：
文献类型：
--
作者：
Bradford,PG;Autieri,M
在人早幼粒细胞白血病细胞 (HL-60) 和用视黄酸 (RA) 或二甲亚砜 (Me2SO) 分化为粒细胞的 HL-60 细胞中检查了 Ins(1,4,5)P3 受体。富含内质网和质膜标记酶活性的 HL-60 细胞膜具有 [3H]Ins(1,4,5)P3 (KD = 22 nM) 的高亲和力结合位点。电转移研究表明 Ins(1,4,[32P]5)P3 与 HL-60 细胞膜的 260 kDa 蛋白质特异性结合。该Ins(1,4,5)P3结合蛋白选择性结合Ca(2+)动员肌醇磷酸盐和其他也与纯化的InsP3受体结合的肌醇磷酸盐，表明HL-60细胞膜的Ins(1,4,5)P3结合蛋白是InsP3受体。当 HL-60 细胞与 1 microM-RA 或 1.25% Me2SO 一起孵育时，细胞在 5-7 天内分化成结构和功能类似于中性粒细胞的细胞。处理后的细胞停止增殖，获得还原硝基蓝四唑染料的能力，并经历分化粒细胞典型的形态变化。随着 HL-60 细胞分化，膜中的最大 [3H]Ins(1,4,5)P3 结合增加 3-4 倍，而 KD 没有变化。结果表明，HL-60细胞分化过程中InsP3受体水平绝对增加，并且这种信号转导蛋白的表达可能受到分化因子的特异性调节。
The Ins(1,4,5)P3 receptor was examined in human promyelocytic leukaemic cells (HL-60) and in HL-60 cells differentiated towards granulocytes with either retinoic acid (RA) or dimethyl sulphoxide (Me2SO). HL-60 cell membranes enriched in marker enzyme activities of the endoplasmic reticulum and the plasma membrane possess a high-affinity binding site for [3H]Ins(1,4,5)P3 (KD = 22 nM). Electrotransfer studies indicate that Ins(1,4,[32P]5)P3 binds specifically to a 260 kDa protein of HL-60 cell membranes. This Ins(1,4,5)P3-binding protein selectively binds Ca(2+)-mobilizing inositol phosphates and other inositol phosphates which also bind to the purified InsP3 receptor, suggesting that the Ins(1,4,5)P3-binding protein of HL-60 cell membranes is the InsP3 receptor. When HL-60 cells are incubated with 1 microM-RA or with 1.25% Me2SO the cells differentiate within 5-7 days into cells resembling neutrophils in both structure and function. Treated cells cease to proliferate, acquire the ability to reduce Nitro Blue Tetrazolium dye, and undergo morphological changes typical of differentiated granulocytes. Concomitant with HL-60 cell differentiation, the maximal [3H]Ins(1,4,5)P3 binding in membranes increases 3-4-fold, with no change in KD. The results suggest that there is an absolute increase in the level of the InsP3 receptor during HL-60 cell differentiation and that the expression of this signal-transducing protein may be specifically regulated by differentiation factors.