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通过荧光显微光解测定单个红细胞血影中的阴离子转运。

Anion transport in single erythrocyte ghosts measured by fluorescence microphotolysis.

作者信息

Peters R, Passow H

出版信息

Biochim Biophys Acta. 1984 Nov 7;777(2):334-8. doi: 10.1016/0005-2736(84)90436-x.

Abstract

Fluorescence microphotolysis was used to measure in single resealed human erythrocyte ghosts the band 3-mediated transport of the fluorescent anion N-(7-nitrobenzofurazan-4-yl)-taurine (NBD-taurine). Transport was reduced to less than 5% of the control by the specific inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS). The accuracy of the determination of the rate constant for NBD-taurine influx was approximately +/- 15% as calculated from repetitive measurements in individual ghosts. The sample population distribution of the rate constant was slightly skewed towards values larger than the mean. The rate of NBD-taurine transport showed an optimum near pH 7. The Arrhenius plot was linear in the range from 28.5 degrees C to 51 degrees C with an apparent activation enthalpy of 21.4 kcal/mol.

摘要

利用荧光显微光解技术,在单个重新封闭的人红细胞血影中测量荧光阴离子N-(7-硝基苯并呋喃唑-4-基)-牛磺酸(NBD-牛磺酸)通过带3介导的转运。特异性抑制剂4,4'-二异硫氰基芪-2,2'-二磺酸盐(DIDS)可使转运减少至对照的5%以下。根据在单个血影中的重复测量计算,NBD-牛磺酸流入速率常数测定的准确度约为±15%。速率常数的样本总体分布略微偏向大于平均值的值。NBD-牛磺酸的转运速率在pH 7附近显示出最佳值。在28.5℃至51℃范围内,阿伦尼乌斯曲线呈线性,表观活化焓为21.4 kcal/mol。

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