Intramitochondrial accumulation of cationic Atto520-biotin proceeds via voltage-dependent slow permeation through lipid membrane
Intramitochondrial accumulation of cationic Atto520-biotin proceeds via voltage-dependent slow permeation through lipid membrane
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DOI：
10.1016/j.bbamem.2015.02.028
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发表时间：
2015-06-01
影响因子：
3.4
通讯作者：
Zorov, Dmitry B.
中科院分区：
文献类型：
--
作者：
Antonenko, Yuri N.;Nechaeva, Natalya L.;Zorov, Dmitry B.
与渗透阳离子的缀合是用于生理活性化合物的线粒体内递送的一般方法，其由在基质侧具有负号的线粒体的高膜电位支持。通过使用荧光相关光谱，我们发现在这里，Atto 520-生物素，共轭物的荧光阳离子罗丹明为基础的染料与膜不可渗透的维生素生物素，积累在通电的线粒体相反，生物素-罗丹明110。Atto 520-生物素的能量依赖性摄取的线粒体，是慢于常规的线粒体染料四甲基罗丹明乙酯，增强了疏水性阴离子四苯硼酸盐（TPB）。Atto 520-生物素也显示出在存在缬氨霉素的情况下由钾离子梯度产生的膜电位驱动的脂质体中蓄积。Atto 520-生物素诱导电流穿过平面双层脂质膜证明了化合物以阳离子形式渗透脂质膜的能力。Atto 520-生物素染色L929细胞培养物中的线粒体，并且在TPB存在下染色增强。因此，荧光Atto 520部分可以作为亲水性药物线粒体内递送的载体。对于生物素-链霉亲和素技术重要的是，发现Atto 520-生物素与链霉亲和素的结合导致其荧光猝灭，类似于荧光素-4-生物素的情况。(C)2015 Elsevier B. V.版权所有。
Conjugation to penetrating cations is a general approach for intramitochondrial delivery of physiologically active compounds, supported by a high membrane potential of mitochondria having negative sign on the matrix side. By using fluorescence correlation spectroscopy, we found here that Atto520-biotin, a conjugate of a fluorescent cationic rhodamine-based dye with the membrane-impermeable vitamin biotin, accumulated in energized mitochondria in contrast to biotin-rhodamine 110. The energy-dependent uptake of Atto520-biotin by mitochondria, being slower than that of the conventional mitochondrial dye tetramethyl-rhodamine ethyl ester, was enhanced by the hydrophobic anion tetraphenylborate (TPB). Atto520-biotin also exhibited accumulation in liposomes driven by membrane potential resulting from potassium ion gradient in the presence valinomycin. The induction of electrical current across planar bilayer lipid membrane by Atto520-biotin proved the ability of the compound to permeate through lipid membrane in a cationic form. Atto520-biotin stained mitochondria in a culture of L929 cells, and the staining was enhanced in the presence of TPB. Therefore, the fluorescent Atto520 moiety can serve as a vehicle for intramitochondrial delivery of hydrophilic drugs. Of importance for biotin-streptavidin technology, binding of Atto520-biotin to streptavidin was found to cause quenching of its fluorescence similar to the case of fluorescein-4-biotin. (C) 2015 Elsevier B.V. All rights reserved.