Costa-Casnellie M R, Segel G B, Lichtman M A
Department of Medicine, University of Rochester Medical Center, New York 14642.
J Biol Chem. 1988 Aug 25;263(24):11851-5.
We have compared the properties of the Na+/H+ exchanger in two cell populations: growing promyelocytic HL-60 cells (immature) and HL-60 cells induced to mature into granulocytes by dimethyl sulfoxide. The exchanger was activated by intracellular acidification from pH 7.25 to pH 5.5. In both immature and mature granulocytic cells, this type of activation resulted in the expected increase in Vmax for Na+ uptake but also in an increase in KmNa. Maximum acidification caused an increase in Vmax of approximately 10-fold in both types of cells. The increase in KmNa was influenced by cell maturation. In immature cells, the KmNa was higher than in mature cells at all pH values tested, and this difference increased with acidification. Maximum acidification increased the KmNa from 15 +/- 4 to 124 +/- 17 mM in immature cells and from 10 +/- 3 to 43 +/- 20 mM in mature cells. Intracellular pH also influenced the pattern of inhibition of 22Na uptake by dimethylamiloride, a specific inhibitor of the exchanger. At intracellular pH 7.0 dimethylamiloride inhibition was mostly competitive in immature and competitive in mature cells. At lower intracellular pH, 5.9, the inhibition was mixed in both types of cells. Thus, the properties of the exchanger in granulocytic cells are influenced by the cell maturation stage and the intracellular pH.
我们比较了两种细胞群体中Na+/H+交换体的特性:正在生长的早幼粒细胞HL-60细胞(未成熟)和经二甲基亚砜诱导成熟为粒细胞的HL-60细胞。该交换体可被细胞内pH从7.25酸化至5.5激活。在未成熟和成熟的粒细胞中,这种类型的激活导致Na+摄取的Vmax预期增加,但同时KmNa也增加。最大酸化导致两种类型细胞的Vmax均增加约10倍。KmNa的增加受细胞成熟的影响。在未成熟细胞中,在所有测试的pH值下,KmNa均高于成熟细胞,并且这种差异随着酸化而增加。最大酸化使未成熟细胞中的KmNa从15±4 mM增加至124±17 mM,使成熟细胞中的KmNa从10±3 mM增加至43±20 mM。细胞内pH也影响了二甲基氨氯吡脒(该交换体的特异性抑制剂)对22Na摄取的抑制模式。在细胞内pH 7.0时,二甲基氨氯吡脒在未成熟细胞中的抑制作用主要为竞争性,在成熟细胞中为竞争性。在较低的细胞内pH 5.9时,两种类型细胞中的抑制作用均为混合型。因此,粒细胞中交换体的特性受细胞成熟阶段和细胞内pH的影响。