Uptake of Cd-Metallothionein into LLC-PK_1 Cells : A Comparative Study with CdCl_2
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概要
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The uptake of the Cd-metallothionein complex (CdMT) into LLC-PK_1 cells was investigated and compared with that of CdCl_2. The cells were incubated at 37℃ for up to 45 min with 1 μM Cd, as either CdMT or CdCl_2 at pH 5.5,6.4,or 7.4. Under all the experimental conditions described below, the accumulation of Cd from CdMT was markedly lower than that from CdCl_2. Cd accumulation at pH 7.4 from CdMT increased linearly with the time of incubation, whereas Cd accumulation from CdCl_2 was saturable. Metabolic inhibitors, 2,4-dinitrophenol(DNP) and carbonylcyanide p-(trifluoromethoxy)-phenylhydrazone (FCCP), and incubation at low temperature significantly decreased Cd accumulation from both Cd compounds. Coincubation with 30 μM ZnCl_2,an antagonist of CdCl_2 uptake, slightly decreased Cd accumulation from CdMT, but it markedly decreased that from CdCl_2. Cd accumulation from CdMT at pH 5.5 was significantly higher than at pH 6.4 or 7.4,whereas Cd accumulation from CdCl_2 at pH 5.5 and 6.4 was significantly lower than at pH 7.4. Although Cd accumulation from CdCl_2 at pH 7.4 was significantly decreased by coincubation with 100 μM cysteine or glutathione (GSH), this decrease was not observed at pH 5.5 or 6.4. A small amount of Cd was removed, by the chelating agent EGTA, from the cell membranes after incubation with CdMT at pH 6.4 and 7.4,whereas a considerable amount of Cd was removed by EGTA after incubation with CdMT at pH 5.5 and with CdCl_2 at three pHs. It appears that the CdMT complex is taken up into LLC-PK_1 cells partially via an energy-dependent process, and the increase in Cd accumulation at low pH is due to the liberation of Cd. High stability and molecular size of the CdMT complex explains why it is not taken up readily into LLC-PK_1 cells.
- 公益社団法人日本薬学会の論文
- 1997-02-15
著者
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Endo T
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Endo Tetsuya
北海道薬科大学
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Sakata M
Department Of Applied Chemistry And Biochemistry Faculty Of Engineering Kumamoto University
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Kimura O
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Endo Tetsuya
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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KIMURA Osamu
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
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SAKATA Masakatsu
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
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Sakata Masakatsu
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Kimura Osamu
Department Of Agricultural Sciences Faculty Of Agriculture Okayama University
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Sakata M
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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