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正常和高血压大鼠主动脉微粒体的能量依赖性钙摄取活性。

Energy-dependent calcium uptake activity of microsomes from the aorta of normal and hypertensive rats.

作者信息

Moore L, Hurwitz L, Davenport G R, Landon E J

出版信息

Biochim Biophys Acta. 1975 Dec 16;413(3):432-43. doi: 10.1016/0005-2736(75)90126-1.

Abstract

Energy-dependent calcium uptake activity of microsomes isolated from the rat aorta has been characterized. The microsomes consist of smooth membrane vesicles which in the presence of MG-ATP as an energy source continuously sequester calcium over a 60-min period. This calcium uptake is greatly stimulated by oxalate anion which serves as a calcium trapping agent. Unlike the calcium uptake of mitochondria this uptake is not inhibited by sodium azide. Sucrose density gradient analysis of the microsomal calcium uptake suggests that the system is associated with the sarcoplasmic reticulum. In presence of 5 mM Mg-ATP and 20 muM calcium approximately 38 nmol of calcium per mg of microsomal protein are taken up in 20 min. In the absence of ATP, less than 2 nmol of calcium per mg of protein are taken up in the first 2 min with no further uptake of calcium in subsequent time periods. When calcium uptake activity is plotted against calcium or ATP concentration of the medium, half maximal activity is calculated for 24.3 muM calcium and for 1.6 mM ATP. The calcium uptake characteristics of the rat aorta microsomes are compatible with a postulated role in the relaxation of the vascular smooth muscle and the provision of an intracellular calcium store for muscle contraction. Aorta microsomes from SHR rats (a genetic strain that is spontaneously hypertensive) have a significantly reduced uptake when compared with the corresponding nonhypertensive control strain. The level of calcium and ATP for half maximal activity of the rat aorta microsomal calcium uptake system is approximately the same in the SHR and the control strain. The rate of release of calcium from rat aorta microsomes is apparently identical in SHR strain and control. The calcium uptake activity of kidney and liver microsomes isolated from the SHR strain and control. The calcium uptake activity of kidney and liver microsomes isolated from the SHR rat appears to be identical to that found in the control strain.

摘要

已对从大鼠主动脉分离出的微粒体的能量依赖性钙摄取活性进行了表征。微粒体由光滑的膜泡组成,在以MG - ATP作为能量来源的情况下,它们在60分钟内持续螯合钙。草酸盐阴离子作为钙捕获剂可极大地刺激这种钙摄取。与线粒体的钙摄取不同,这种摄取不受叠氮化钠的抑制。对微粒体钙摄取的蔗糖密度梯度分析表明,该系统与肌浆网相关。在存在5 mM Mg - ATP和20 μM钙的情况下,每毫克微粒体蛋白在20分钟内大约摄取38 nmol钙。在没有ATP的情况下,每毫克蛋白在前2分钟内摄取的钙少于2 nmol,在随后的时间段内没有进一步的钙摄取。当将钙摄取活性与培养基中的钙或ATP浓度作图时,计算得出钙浓度为24.3 μM和ATP浓度为1.6 mM时的半数最大活性。大鼠主动脉微粒体的钙摄取特征与推测的在血管平滑肌舒张以及为肌肉收缩提供细胞内钙库中的作用相符。与相应的非高血压对照品系相比,SHR大鼠(一种遗传性自发性高血压品系)的主动脉微粒体摄取明显减少。SHR大鼠和对照品系中,大鼠主动脉微粒体钙摄取系统半数最大活性时的钙和ATP水平大致相同。SHR品系和对照品系中,从大鼠主动脉微粒体释放钙的速率显然相同。从SHR大鼠和对照中分离出的肾脏和肝脏微粒体的钙摄取活性。从SHR大鼠分离出的肾脏和肝脏微粒体的钙摄取活性似乎与对照品系中的相同。

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