Fry A J, Towner P, Holman G D, Eisenthal R
Department of Biochemistry, University of Bath, UK.
Mol Biochem Parasitol. 1993 Jul;60(1):9-18. doi: 10.1016/0166-6851(93)90023-q.
Kinetic parameters for entry of D-fructose into Trypanosoma brucei brucei have been determined. The net uptake of D-fructose was found to be rapid and occurred at a rate which was comparable with that observed for uptake of D-glucose. The Km and Vmax were 3.91 +/- 1.58 mM and 69.1 +/- 7.2 nmol min-1 (mg protein)-1. D-Fructose was metabolized to pyruvate under aerobic conditions and to pyruvate and glycerol under anaerobic conditions in a manner similar to D-glucose. Comparisons of the kinetic parameters for D-fructose transport and metabolism indicated that uptake was rate limiting. Inhibition constants (Ki) for inhibition of 6-deoxy-D-glucose by D-fructose and D-fructose transport by 6-deoxy-D-glucose were consistent with the Km values for these two substrates. These interactions indicate that D-fructose and 6-deoxy-D-glucose share a single common transporter. 1,5-Anhydro-D-glucitol and 1,5 anhydro-D-mannitol (the fused pyranose ring analogues of D-glucose and D-mannose) have been found to interact well with the transporter, while L-sorbose (a D-fructose analogue with a pyranose ring) had only low affinity. However, 2,5-anhydro-D-mannitol (a fused furanose ring analogue of D-fructose) inhibited both 6-deoxy-D-glucose and D-fructose transport with a Ki of approx. 0.8 mM. The high affinity for 2,5-anhydro-D-mannitol (2-deoxy-D-fructofuranose) indicates that D-fructose is transported in the furanose ring form.(ABSTRACT TRUNCATED AT 250 WORDS)
已测定D-果糖进入布氏布氏锥虫的动力学参数。发现D-果糖的净摄取迅速,摄取速率与D-葡萄糖摄取速率相当。米氏常数(Km)和最大反应速度(Vmax)分别为3.91±1.58 mM和69.1±7.2 nmol·min⁻¹·(mg蛋白质)⁻¹。在有氧条件下,D-果糖代谢为丙酮酸,在无氧条件下代谢为丙酮酸和甘油,其方式与D-葡萄糖相似。D-果糖转运和代谢的动力学参数比较表明,摄取是限速步骤。D-果糖对6-脱氧-D-葡萄糖摄取的抑制常数(Ki)以及6-脱氧-D-葡萄糖对D-果糖转运的抑制常数与这两种底物的Km值一致。这些相互作用表明D-果糖和6-脱氧-D-葡萄糖共享一个共同的转运体。已发现1,5-脱水-D-葡萄糖醇和1,5-脱水-D-甘露糖醇(D-葡萄糖和D-甘露糖的稠合吡喃糖环类似物)与该转运体相互作用良好,而L-山梨糖(具有吡喃糖环的D-果糖类似物)亲和力较低。然而,2,5-脱水-D-甘露糖醇(D-果糖的稠合呋喃糖环类似物)以约0.8 mM的Ki抑制6-脱氧-D-葡萄糖和D-果糖的转运。对2,5-脱水-D-甘露糖醇(2-脱氧-D-果糖呋喃糖)的高亲和力表明D-果糖以呋喃糖环形式转运。(摘要截断于250字)