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大鼠肝脏溶酶体的中性糖转运

Neutral-sugar transport by rat liver lysosomes.

作者信息

Jonas A J, Conrad P, Jobe H

机构信息

Division of Medical Genetics, Harbor-UCLA Medical Center, Torrance 90502.

出版信息

Biochem J. 1990 Dec 1;272(2):323-6. doi: 10.1042/bj2720323.

Abstract

Transport of D-glucose was studied in Percoll-gradient-purified rat liver lysosomes. D-Glucose uptake had a Km of 22 mM and a t1/2 of approx. 30 s. D-Fucose, 2-deoxyglucose and methyl alpha-glucoside were the most effective competitors for uptake of D-glucose, although D-galactose, D-mannose, D-xylose and L-fucose also appeared to compete for uptake. L-Glucose was a poor competitor for uptake. No competition was observed with N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-glucuronic acid, N-acetylneuraminic acid, D-glucosamine or the amino acids L-glycine, L-lysine and L-proline. Uptake was unaffected by N-ethylmaleimide, dithiothreitol, KCl, NaCl, ATP/Mg or alteration of buffer pH. D-Glucose efflux from lysosomes was temperature-dependent, with a Q10 of 2.3, and was inhibited by cytochalasin B. Counter-transport could not be demonstrated. In contrast, L-fucose uptake had a Km of 65 mM and was largely unaffected by 5 M excess of neutral D-sugars. Both uptake and efflux of L-fucose were inhibited by cytochalasin B. It appears that lysosomes possess a facilitated transport system for D-glucose and perhaps other neutral D-sugars that is discrete from transport systems for acetylated and acidic sugars.

摘要

在经Percoll梯度纯化的大鼠肝脏溶酶体中研究了D-葡萄糖的转运。D-葡萄糖摄取的Km为22 mM,t1/2约为30秒。D-岩藻糖、2-脱氧葡萄糖和α-甲基葡萄糖苷是D-葡萄糖摄取的最有效竞争者,尽管D-半乳糖、D-甘露糖、D-木糖和L-岩藻糖似乎也参与竞争摄取。L-葡萄糖是摄取的弱竞争者。未观察到与N-乙酰-D-葡萄糖胺、N-乙酰-D-半乳糖胺、D-葡萄糖醛酸、N-乙酰神经氨酸、D-葡萄糖胺或氨基酸L-甘氨酸、L-赖氨酸和L-脯氨酸存在竞争。摄取不受N-乙基马来酰亚胺、二硫苏糖醇、KCl、NaCl、ATP/Mg或缓冲液pH改变的影响。溶酶体中D-葡萄糖的外排是温度依赖性的,Q10为2.3,并受细胞松弛素B抑制。未证实存在反向转运。相比之下,L-岩藻糖摄取的Km为65 mM,并且在5 M过量的中性D-糖存在时基本不受影响。L-岩藻糖的摄取和外排均受细胞松弛素B抑制。看来溶酶体拥有一种促进D-葡萄糖以及可能其他中性D-糖转运的系统,该系统与乙酰化糖和酸性糖的转运系统不同。

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THIN-LAYER CHROMATOGRAPHY ON MICROCRYSTALLINE CELLULOSE.微晶纤维素上的薄层色谱法
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