MITOCHONDRIAL H+ CO-TRANSPORTERS: GLUTAMATE AND PYRUVATE

线粒体内膜的几个阴离子转运系统出现 作为质子补偿的电中性转运蛋白。 是 这项研究的目的是确定蛋白质的内部 大鼠脑和肝脏、牛心和绿头苍蝇的线粒体膜 来自发育和衰老动物的飞行肌， 丙酮酸转运和谷氨酸转运，并确定功能 与磷酸盐不同或相同的参数 转运蛋白 丙酮酸转运系统（单羧酸 转运蛋白，质子补偿，电中性）似乎存在于 所有研究的线粒体，即使它可能发生在不同的最大值 来自不同器官的线粒体中的周转率或浓度， 不同的生物体。 谷氨酸转运活动已经 在某些类型的线粒体中不存在， 其他线粒体中的不同活性可能是由于 比活性或转运蛋白浓度。 谷氨酸 转运蛋白也是质子补偿和电中性的。 我们计划 鉴定十二烷基硫酸盐聚丙烯酰胺凝胶中的转运蛋白 在用放射性SH标记的基础上电泳后， 试剂 虽然识别标记的蛋白质很重要，但 必须在染色凝胶（银或考马斯）上检测蛋白质 蓝色染色），以确定其在各种级分中的存在 在净化阶段。 我们希望能够使用我们的新 修改重构程序以将蛋白质掺入活性物质中， 形成脂质体以详细研究其性质。 线粒体对细胞代谢至关重要， 代谢阴离子必须被转运到线粒体基质中。 的 仅阴离子转运系统本身不是代谢物转运蛋白 但我们开始了解腺嘌呤 核苷酸移位酶（产电阴离子交换转运蛋白）和 磷酸盐转运蛋白（质子补偿电中性转运蛋白）。 它 因此，对确定运输和确定其特点具有根本重要性 负责阴离子代谢物转运的蛋白质。

Several anion transport systems of the inner mitrochondrial membrane appear to function as proton compensated electroneutral transporters. It is the objective of this research proposal to identify the proteins of the inner mitochondrial membrane from rat brain and liver, beef heart, and blowfly flight muscle from developing and senescent animals that are responsible for pyruvate transport and glutamate transport and identify functional parameters that are different from or the same as those of the phosphate transport protein. The pyruvate transport system (monocaboxylate transporter, proton compensated, electroneutral) appears to be present in all mitochondria studied even though it may occur with different maximum turnover rates or concentrations in mitochondria from different organs or different organisms. The glutamate transport activity has already been demonstrated to be absent from some types of mitochondria while again different activities in other mitochondria may be due to differences in specific activities or transport protein concentrations. The glutamate transporter also is proton compensated and electroneutral. We plan to identify the transport proteins in dodecyl sulfate polyacrylamide gels after electrophoresis on the basis of labeling with radioactive SH reagents. While it is important to identify the labeled protein, it is necessary that the protein is detected on stained gels (Silver or Coomassie Blue stain) in order to determine its presence in the various fractions during stages of purification. We expect to be able to use our newly modified reconstitution procedure to incorporate the proteins in an active form into liposomes in order to study their properties in detail. Mitochondria are of critical importance to the cellular metabolism and metabolic anions have to be transported into the mitochondrial matrix. The only anion transport system which are not metabolite transporters as such but for which we are beginning to develop an understanding are the adenine nucleotide translocase (electrogenic anion exchange transporter) and the phosphate transporter (proton compensated electroneutral transporter). It is thus of fundamental importance to identify and characterize transport proteins that are responsible for anion metabolite transport.