Kato M, Yang J, Iwai T, Tanamura A, Arino T, Kawashima O, Takeda N
Department of Internal Medicine, Aoto Hospital, Jikei University School of Medicine, Tokyo, Japan.
Mol Cell Biochem. 1993 Feb 17;119(1-2):89-94. doi: 10.1007/BF00926858.
ADP/ATP carrier protein (AAC) is located in the mitochondrial inner membrane and has an important function in mitochondrial energy supply. This protein transports ATP to the cytoplasm and counter transports ADP into the mitochondria. J-2-N cardiomyopathic hamsters were investigated to determine the AAC content in cardiac mitochondria. After recording an electrocardiogram and collecting blood, the cardiac mitochondria were isolated. The mitochondrial membranes were labelled with eosin-5-maleimide (EMA) and separated on SDS polyacrylamide gels. The position of the AAC component was identified by exposing the gel under UV light, and the AAC content was determined by densitometry after staining with Coomassie blue. The AAC content ratio was significantly decreased in both 10-week-old and 1-year survived J-2-N hamsters when compared to control Golden hamster. Among 10-week-old J-2-N hamsters, the decrease in the AAC content ratio was more marked for the animals with more severe myocardial damage. The H(+)-ATPase activities of mitochondrial membrane were higher in 10-week-old J-2-N hamsters than in control hamsters. These results suggest that the decrease of AAC in J-2-N hamster plays an important role in the pathogenesis of cardiomyopathy in J-2-N hamsters.
ADP/ATP载体蛋白(AAC)位于线粒体内膜,在线粒体能量供应中发挥重要作用。该蛋白将ATP转运至细胞质,并将ADP反向转运至线粒体。对J-2-N心肌病仓鼠进行研究,以确定心脏线粒体中的AAC含量。记录心电图并采集血液后,分离出心脏线粒体。线粒体膜用马来酰亚胺基-5-异硫氰酸荧光素(EMA)标记,并在SDS聚丙烯酰胺凝胶上进行分离。通过在紫外光下曝光凝胶来鉴定AAC成分的位置,并用考马斯亮蓝染色后通过光密度测定法确定AAC含量。与对照金黄仓鼠相比,10周龄和存活1年的J-2-N仓鼠的AAC含量比率均显著降低。在10周龄的J-2-N仓鼠中,心肌损伤越严重的动物,AAC含量比率下降越明显。10周龄J-2-N仓鼠线粒体膜的H(+)-ATP酶活性高于对照仓鼠。这些结果表明,J-2-N仓鼠中AAC的减少在J-2-N仓鼠心肌病的发病机制中起重要作用。