Kako K J, Patterson S D
Biochem J. 1975 Nov;152(2):313-23. doi: 10.1042/bj1520313.
Activities of phosphatidate phosphohydrolase and palmitoyl-CoA hydrolase were determined in cardiac subcellular fractions prepared from rabbits which has received tri-iodothyronine and from hamsters with hereditary cardiomyopathy (strain BIO 14.6). 1. Both mitochondrial and microsomal fractions of hyperthyroid rabbit hearts produced 4-5 times as much diacylglycerol 3-phosphate from glycerol 3-phosphate and palmitate as did those of euthyroid hearts. 2. Phosphatidate phosphohydrolase, measured with phosphatidate emulsion, was activated by 1mm-Mg(2+) in all but the mitochondrial fraction of euthyroid rabbit hearts. The activation was more pronounced in subcellular fractions isolated from hyperthyroid hearts, so that the measured activities were significantly increased above those of the controls. The highest activity was found in the microsomal and lysosomal fractions. 3. In the absence of Mg(2+) during incubation, the difference in phosphohydrolase activities between eu- and hyper-thyroid states was not significant. 4. The phosphohydrolase of subcellular fractions of control hamsters did not respond to addition of 0.5-8.0mm-Mg(2+). The enzyme from cardiomyopathic hearts was slightly inhibited by this bivalent cation and therefore significant increases in activity were observed only in the absence of Mg(2+) from the assay system. 5. The rate of reaction by soluble phosphatidate phosphohydrolase was similar regardless of the nature of the substrate. Both when microsomal-bound phosphatidate was used as the substrate and when phosphatidate suspension was used, the activity of soluble enzyme was lower than that of the microsomal and lysosomal enzymes measured with phosphatidate suspension; this was especially so when the assay was carried out in the absence of Mg(2+). Neither tri-iodothyronine nor cardiomyopathy influenced the soluble phosphohydrolase activity in the two species. 6. Neither tri-iodothyronine nor cardiomyopathy significantly changed palmitoyl-CoA hydrolase activities in subcellular fractions. 7. Microsomal diacylglycerol acyltransferase and myocardial triacylglycerol content were also unchanged in the hyperthyroid state.
在从接受过三碘甲状腺原氨酸的兔子以及患有遗传性心肌病(BIO 14.6品系)的仓鼠制备的心脏亚细胞组分中,测定了磷脂酸磷酸水解酶和棕榈酰辅酶A水解酶的活性。1. 甲状腺功能亢进的兔子心脏的线粒体和微粒体组分从甘油3 - 磷酸和棕榈酸产生的3 - 磷酸二酰甘油是甲状腺功能正常的心脏的4 - 5倍。2. 用磷脂酸乳剂测定时,除甲状腺功能正常的兔子心脏的线粒体组分外,所有组分中的磷脂酸磷酸水解酶都被1mM的Mg(2+)激活。在从甲状腺功能亢进的心脏分离的亚细胞组分中,这种激活更为明显,因此测得的活性比对照组显著增加。最高活性出现在微粒体和溶酶体组分中。3. 在孵育过程中不存在Mg(2+)时,甲状腺功能正常和亢进状态之间的磷酸水解酶活性差异不显著。4. 对照仓鼠亚细胞组分的磷酸水解酶对添加0.5 - 8.0mM的Mg(2+)没有反应。来自心肌病心脏的酶被这种二价阳离子轻微抑制,因此仅在测定系统中不存在Mg(2+)时才观察到活性显著增加。5. 可溶性磷脂酸磷酸水解酶的反应速率与底物的性质无关。当使用微粒体结合的磷脂酸作为底物以及使用磷脂酸悬浮液时,可溶性酶的活性低于用磷脂酸悬浮液测定的微粒体和溶酶体酶的活性;在不存在Mg(2+)进行测定时尤其如此。三碘甲状腺原氨酸和心肌病都不影响这两个物种中的可溶性磷酸水解酶活性。6. 三碘甲状腺原氨酸和心肌病都没有显著改变亚细胞组分中棕榈酰辅酶A水解酶的活性。7. 在甲状腺功能亢进状态下,微粒体二酰甘油酰基转移酶和心肌三酰甘油含量也没有变化。