Grubbs R D, Collins S D, Maguire M E
J Biol Chem. 1984 Oct 10;259(19):12184-92.
28Mg2+ influx studies in S49 murine lymphoma cells indicate that only 2-3% of total cell Mg2+ content can be exchanged at isotopic equilibrium, implying compartmentation of the newly transported Mg2+. The nature of this compartmentation was examined using selective permeabilization of the plasma membrane with the detergent, digitonin. Control experiments demonstrated that the digitonin permeabilization procedure did not release mitochondrial and lysosomal components, alter mitochondrial respiration, or significantly change cell morphology. Thus, under appropriate conditions, the digitonin permeabilization technique allows determination of the amount of a particular cell constituent within the solute space of the cytoplasm. In nonproliferating cells at an extracellular Mg2+ concentration of 0.1 mM, newly transported Mg2+ equilibrates within 2 h with a small cytoplasmic Mg2+ pool comprising about 3% of the total cytoplasmic Mg2+ (about 2% of total cell Mg2+). The pool of Mg2+ does not equilibrate with bulk cytoplasmic or cellular Mg2+ for at least 16 h. The Mg2+ pool size is dependent on extra-cellular Mg2+ concentration, is saturable with increasing extracellular Mg2+, and reaches a maximal size of 6-7% of total cell Mg2+ at 2 mM extracellular Mg2+. Unlike Mg2+, newly transported Ca2+ is quickly sequestered in noncytoplasmic compartments. In proliferating cells, however, newly transported Mg2+ exchanges extensively with cytoplasmic Mg2+ over the course of 4 h, suggesting that compartmentation of Mg2+ may be dependent on proliferative status.
对S49鼠淋巴瘤细胞中28Mg2+内流的研究表明,在同位素平衡时,细胞内总Mg2+含量中只有2 - 3%能够交换,这意味着新转运的Mg2+存在区室化现象。利用去污剂洋地黄皂苷对质膜进行选择性通透处理,研究了这种区室化的性质。对照实验表明,洋地黄皂苷通透处理过程不会释放线粒体和溶酶体成分,不会改变线粒体呼吸,也不会显著改变细胞形态。因此,在适当条件下,洋地黄皂苷通透技术可以测定细胞质溶质空间内特定细胞成分的含量。在细胞外Mg2+浓度为0.1 mM的非增殖细胞中,新转运的Mg2+在2小时内与一个小的细胞质Mg2+池达到平衡,该池约占细胞质总Mg2+的3%(约占细胞总Mg2+的2%)。Mg2+池至少16小时内不会与大量细胞质或细胞内Mg2+达到平衡。Mg2+池大小取决于细胞外Mg2+浓度,随着细胞外Mg2+浓度增加而饱和,在细胞外Mg2+浓度为2 mM时达到细胞总Mg2+的6 - 7%的最大大小。与Mg2+不同,新转运的Ca2+很快被隔离在非细胞质区室中。然而,在增殖细胞中,新转运的Mg2+在4小时内与细胞质Mg2+广泛交换,这表明Mg2+的区室化可能取决于增殖状态。