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巯基试剂和光亲和探针对“L”和“Ly+”氨基酸转运系统的差异性失活作用

Differential inactivation of the "L" and "Ly+" amino acid transport systems by a sulfhydryl reagent and a photo-affinity probe.

作者信息

Hare J D, Marinetti G V, Meisler A I, Tometsko A M

出版信息

Biochim Biophys Acta. 1976 Sep 7;443(3):485-93. doi: 10.1016/0005-2736(76)90467-3.

Abstract

Using a substrate-stimulated amino acid efflux system, it has been shown that the "Ly+" and "L" amino acid transport systems of mouse embryo cells in culture are differentially inhibited by parachloromercuribenzene sulfonate (PCMB-S) and the photoaffinity probe 4-fluoro-3-nitrophenylazide (FNPA). Three types of evidence support the conclusion that these transport systems are mediated by separate carrier proteins. (1) The specificity of substrate-stimulated efflux is high for each system; (2) PCMB-S inhibits L-phenylalanine and L-leucine stimulated L-[3H]phenylalanine efflux with no effect on L-lysine stimulated L-[3H]lysine efflux, and (3) the photo-affinity probe FNPA inhibits L-lysine efflux with little effect on the L-phenylalanine-stimulated efflux.

摘要

利用底物刺激的氨基酸外排系统,已表明培养的小鼠胚胎细胞的“Ly+”和“L”氨基酸转运系统受到对氯汞苯磺酸盐(PCMB-S)和光亲和探针4-氟-3-硝基苯叠氮化物(FNPA)的不同抑制。三类证据支持这些转运系统由不同载体蛋白介导的结论。(1)每个系统的底物刺激外排特异性都很高;(2)PCMB-S抑制L-苯丙氨酸和L-亮氨酸刺激的L-[3H]苯丙氨酸外排,而对L-赖氨酸刺激的L-[3H]赖氨酸外排无影响;(3)光亲和探针FNPA抑制L-赖氨酸外排,对L-苯丙氨酸刺激的外排影响很小。

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