Henslee J G, Srere P A
J Biol Chem. 1979 Jun 25;254(12):5488-97.
The mitochondrial matrix subfractions from rat liver, kidney cortex, brain, heart, and skeletal muscle were isolated and their protein components were resolved by two-dimensional polyacrylamide gel electrophoresis, revealing between 120 and 150 components for each matrix subfraction. Excellent resolution was obtained utilizing a pH 5 to 8 gradient in the first dimension and in 8 to 13% exponential acrylamide gradient in the second dimension, increasing the number of mitochondrial matrix proteins observed 3-fold over one-dimensional systems. Protein components tentatively identified by co-migration with pure enzymes and by known tissue distributions are carbamoyl-phosphate synthetase (EC 2.7.2.5), ornithine transcarbamylase (EC 2.1.3.3), glutamate dehydrogenase (EC 1.4.1.3), pyruvate carboxylase (EC 6.4.1.1), citrate synthase (EC 4.1.3.7), fumarase (EC 4.2.1.2), aconitase (EC 4.2.1.3), alpha-ketoglutarate dehydrogenase (EC 1.2.4.2), dihydrolipoyl transsuccinylase (EC 2.3.1.12), lipoamide dehydrogenase (EC 1.6.4.3), glutamate-aspartate aminotransferase (EC 2.6.1.1), and the two subunits of pyruvate dehydrogenase (EC 1.2.4.1). Protein components unambiguously identified by peptide mapping are citrate synthase, aconitase, and pyruvate carboxylase. The inner membrane subfraction from rat liver mitochondria was also resolved two dimensionally; the alpha and beta subunits of ATPase (F1) (EC 3.6.1.3) were identified by peptide mapping.
分离出大鼠肝脏、肾皮质、脑、心脏和骨骼肌的线粒体基质亚组分，通过二维聚丙烯酰胺凝胶电泳解析其蛋白质成分，结果显示每个基质亚组分含有120至150种成分。在第一维使用pH 5至8的梯度，在第二维使用8%至13%的指数丙烯酰胺梯度，获得了极佳的分辨率，与一维系统相比，观察到的线粒体基质蛋白数量增加了3倍。通过与纯酶共迁移以及已知组织分布初步鉴定的蛋白质成分有氨甲酰磷酸合成酶（EC 2.7.2.5）、鸟氨酸转氨甲酰酶（EC 2.1.3.3）、谷氨酸脱氢酶（EC 1.4.1.3）、丙酮酸羧化酶（EC 6.4.1.1）、柠檬酸合酶（EC 4.1.3.7）、延胡索酸酶（EC 4.2.1.2）、乌头酸酶（EC 4.2.1.3）、α-酮戊二酸脱氢酶（EC 1.2.4.2）、二氢硫辛酰转琥珀酰酶（EC 2.3.1.12）、硫辛酰胺脱氢酶（EC 1.6.4.3）、谷氨酸-天冬氨酸转氨酶（EC 2.6.1.1）以及丙酮酸脱氢酶（EC 1.2.4.1）的两个亚基。通过肽图谱明确鉴定的蛋白质成分有柠檬酸合酶、乌头酸酶和丙酮酸羧化酶。还对大鼠肝脏线粒体的内膜亚组分进行了二维解析；通过肽图谱鉴定出了ATP酶（F1）（EC 3.6.1.3）的α和β亚基。
