Evaluation method for polyamine uptake by N(1)-dansylspermine
Evaluation method for polyamine uptake by N(1)-dansylspermine
复制标题
N(1)-丹酰精胺摄取多胺的评价方法
DOI：
10.1007/s00726-009-0414-9
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Akira Shirahata
中科院分区：
文献类型：
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作者：
Koichi Takao;Yoshiaki Sugita;Akira Shirahata
Polyamine uptake by the polyamine transport system (PTS) in HTC cells was studied without the use of radioisotope-labeled polyamines. N1-Dansylspermine (DNS343) was selected as a candidate probe to examine the PTS. DNS343 was incorporated into HTC cells, and its distribution in the cells was confirmed by fluorescence microscopy. The incorporation of DNS343 via PTS was confirmed by a competition study with bis(3-aminopropyl)amine, which is incorporated into cells via the PTS. In addition, the temperature dependency of DNS343 uptake and studies with inhibitors of ornithine decarboxylase and proteoglycan synthesis supported the use of DNS343 as a fluorescent probe for the PTS. The kinetics studies for HTC cells treated with or without an ornithine decarboxylase inhibitor indicated that DNS343 uptake was saturable and that the apparent Km values for the PTS were approximately 1.5 μM in both types of cells at 37°C. Thus, we developed an assay method for the PTS by high-performance liquid-chromatography with DNS343. The inhibitory effect of polyamine analogs and related compounds on DNS343 uptake was then examined and discussed.
