Yamanouchi T, Tachibana Y, Sekino N, Akanuma H, Akaoka I, Miyashita H
Second Department of Internal Medicine, University of Teikyo, Tokyo, Japan.
J Biol Chem. 1994 Apr 1;269(13):9664-8.
The transport and intracellular accumulation of 1,5-anhydro-D-glucitol (AG) was studied in the human erythroleukemia cell line K-562 by gas chromatography-mass spectrometry in conjunction with liquid scintillation spectrometry. K-562 cells contained 106 +/- 6 nM/10(6) cells of free AG, primarily in the cytosol. Addition of physiologic amounts of AG to the extracellular medium resulted in rapid intracellular incorporation of AG, with a half-saturation time of 5 s. Intracellular accumulation was linear for 2 h and subsequently reached saturation. AG uptake was temperature and concentration dependent with an apparent Km of 127 mM. AG uptake and accumulation was not inhibited by fructose, fucose, galactose, mannose, glucose, or 3-O-methyl-D-glucose and was less affected by cytochalasin B or phloretin than that of 2-deoxyglucose. Phloridzin did not affect AG uptake but did inhibit 2-deoxyglucose uptake. Efflux of AG from K-562 cells depended on external AG concentration alone and was not affected by extracellular glucose concentration. Intracellular AG concentration decreased rapidly and reached zero within 10 min following removal of AG from the external medium. We therefore propose that both transport and countertransport of AG in K-562 cells are mediated by a specific carrier system.
采用气相色谱-质谱联用技术结合液体闪烁光谱法,对人红白血病细胞系K-562中1,5-脱水-D-葡萄糖醇(AG)的转运和细胞内蓄积进行了研究。K-562细胞含有106±6 nM/10(6)个细胞的游离AG,主要存在于细胞质中。向细胞外培养基中添加生理量的AG会导致AG迅速在细胞内掺入,半饱和时间为5秒。细胞内蓄积在2小时内呈线性,随后达到饱和。AG摄取具有温度和浓度依赖性,表观Km为127 mM。果糖、岩藻糖、半乳糖、甘露糖、葡萄糖或3-O-甲基-D-葡萄糖均不抑制AG摄取和蓄积,与2-脱氧葡萄糖相比,细胞松弛素B或根皮素对其影响较小。根皮苷不影响AG摄取,但抑制2-脱氧葡萄糖摄取。AG从K-562细胞的流出仅取决于外部AG浓度,不受细胞外葡萄糖浓度影响。从外部培养基中去除AG后,细胞内AG浓度迅速下降,并在10分钟内降至零。因此,我们认为K-562细胞中AG的转运和逆向转运均由特定的载体系统介导。