Brailsford M A, Thompson A G, Kaderbhai N, Beechey R B
Biochem Biophys Res Commun. 1986 Nov 14;140(3):1036-42. doi: 10.1016/0006-291x(86)90739-4.
The pyruvate carrier from castor bean mitochondria has been solubilized with Triton X-114 and partially purified using hydroxyapatite column chromatography. SDS-polyacrylamide gel electrophoresis of the hydroxyapatite-eluate showed that there were 6 major protein bands of Mr, 74kDa, 66kDa, 34kDa, 32kDa, 30kDa 12kDa. When the eluate was reconstituted into liposomes it was shown to catalyze a pyruvate exchange reaction which was sensitive to N-ethyl maleimide and a series of analogues of alpha-cyanocinnamate. The characteristics of this pyruvate exchange activity are similar to that found in intact mitochondria, and it is concluded that one or more proteins in the hydroxyapatite-eluate correspond to the pyruvate carrier.
蓖麻籽线粒体中的丙酮酸载体已用Triton X-114增溶，并通过羟基磷灰石柱色谱法进行了部分纯化。羟基磷灰石洗脱液的SDS-聚丙烯酰胺凝胶电泳显示，有6条主要蛋白带，分子量分别为74kDa、66kDa、34kDa、32kDa、30kDa和12kDa。当洗脱液重构到脂质体中时，它被证明能催化丙酮酸交换反应，该反应对N-乙基马来酰亚胺和一系列α-氰基肉桂酸类似物敏感。这种丙酮酸交换活性的特征与完整线粒体中的相似，因此得出结论，羟基磷灰石洗脱液中的一种或多种蛋白质对应于丙酮酸载体。
