Wiemer E A, Ter Kuile B H, Michels P A, Opperdoes F R
International Institute of Cellular and Molecular Pathology, Research Unit for Tropical Diseases, Brussels, Belgium.
Biochem Biophys Res Commun. 1992 Apr 30;184(2):1028-34. doi: 10.1016/0006-291x(92)90694-g.
The characteristics of pyruvate transport across the plasma membrane in the bloodstream form of Trypanosoma brucei were studied using [14C]pyruvate in combination with the silicone-oil centrifugation technique. We present evidence for the existence of a facilitated diffusion carrier in the plasma membrane of T. brucei which specifically mediates the translocation of pyruvate. The uptake of pyruvate followed saturation kinetics (Km 1.96 +/- 0.28 mM; Cmax 36.61 +/- 1.15 nmol pyruvate/30 sec.mg protein), after correction of the data for a nonsaturable diffusion component. The uptake of pyruvate was competitively inhibited by a number of (oxo)monocarboxylic acids, including pyruvate analogs and metabolically related substances, but not by L-lactate. The transport exhibited the phenomenon of transacceleration, indicative for the involvement of a facilitated diffusion carrier. The carrier is highly specific for pyruvate and differs from other known monocarboxylate carriers present in the mitochondrial and/or plasma membrane of other eukaryotic cells in that it does not transport L-lactate.
利用[14C]丙酮酸结合硅油离心技术,研究了布氏锥虫血流形式下丙酮酸跨质膜转运的特性。我们提供证据表明,布氏锥虫质膜中存在一种促进扩散载体,它特异性介导丙酮酸的转运。校正非饱和扩散成分的数据后,丙酮酸的摄取遵循饱和动力学(Km 1.96 +/- 0.28 mM;Cmax 36.61 +/- 1.15 nmol丙酮酸/30秒·毫克蛋白)。丙酮酸的摄取受到多种(氧代)单羧酸的竞争性抑制,包括丙酮酸类似物和代谢相关物质,但不受L-乳酸的抑制。转运表现出转加速现象,表明存在促进扩散载体。该载体对丙酮酸具有高度特异性,与其他真核细胞线粒体和/或质膜中存在的其他已知单羧酸载体不同,它不转运L-乳酸。