Veklich T O, Kosterin S O, Shynlova O P
Palladin Institute of Biochemistry, NAS of Ukraine, Kyiv.
Ukr Biokhim Zh (1999). 2002 Jan-Feb;74(1):42-8.
In the experiments conducted with application of an isotopic technique (45Ca2+) on the myometrium cells suspension treated by digitonin solution (0.1 mg/ml) some properties of Ca ions accumulation system in the mitochondria--cationic and substrate specificity as well as effects of Mg2+ and some other bivalent metals ions on the Ca2+ accumulation velocity have been estimated. Ca ions accumulation from the incubation medium containing 3 mM sodium succinate Na, 2 mM Pi (as potassium K(+)-phosphate buffer, pH 7.4 at 37 degrees C), 0.01 mM (40CaCl2 + 45CaCl2) and 100 nM thapsigargin--selective inhibiting agent of endoplasmatic reticulum calcium pump were demonstrated as detected just only in presence of Mg, while not Ni, Co or Cu ions. The increase of Mg2+ concentration from 1 x 10(-6) to 10(-3) M induced the ATP dependent transport activation in the myometrium mitochondria. Under [Mg2+] increase till 40 mM this cation essentially decreased Ca2+ accumulation (by 65% from the maximal value). The optimum for Ca2+ transport in the myometrium cells suspension is Mg2+ 10 mM concentration. Ka activation apparent constant along Mg2+ value (in presence 3 mM ATP and 3 mM sodium succinate) is 4.27 mM. The above listed bivalent metals decreased Mg2+, ATP-dependent accumulation of calcium, values of inhibition apparent constants for ions Co2+, Ni2+ and Cu2+ were--2.9 x 10(-4) M, 5.1 x 10(-5) M and 4.2 x 10(-6) M respectively. For Mg2+, ATP-dependent Ca2+ transport in the uterus myocytes mitocondria a high substrate specificity is a characteristic phenomenon in elation to ATP: GTP, CTP and UTP practically fail to provide for Ca accumulation process.
在用同位素技术(45Ca2+)对经洋地黄皂苷溶液(0.1mg/ml)处理的子宫肌层细胞悬液进行的实验中,评估了线粒体中钙离子积累系统的一些特性——阳离子和底物特异性,以及Mg2+和其他一些二价金属离子对Ca2+积累速度的影响。在含有3mM琥珀酸钠Na、2mM Pi(作为钾K(+)-磷酸盐缓冲液,37℃时pH 7.4)、0.01mM(40CaCl2 + 45CaCl2)和100nM毒胡萝卜素(内质网钙泵的选择性抑制剂)的孵育培养基中,仅在存在Mg而不存在Ni、Co或Cu离子的情况下,才检测到Ca离子的积累。Mg2+浓度从1×10(-6) M增加到10(-3) M会诱导子宫肌层线粒体中ATP依赖性转运激活。当[Mg2+]增加到40mM时,这种阳离子会显著降低Ca2+积累(从最大值降低65%)。子宫肌层细胞悬液中Ca2+转运的最佳Mg2+浓度为10mM。沿着Mg2+值(在存在3mM ATP和3mM琥珀酸钠的情况下)的Ka激活表观常数为4.27mM。上述二价金属降低了Mg2+、ATP依赖性的钙积累,离子Co2+、Ni2+和Cu2+的抑制表观常数分别为——2.9×10(-4) M、5.1×10(-5) M和4.2×10(-6) M。对于子宫肌细胞线粒体中Mg2+、ATP依赖性的Ca2+转运,相对于ATP具有高底物特异性是一个特征现象:GTP、CTP和UTP实际上无法支持Ca积累过程。