Restoration of mutant bestrophin-1 expression, localisation and function in a polarised epithelial cell model.
Restoration of mutant bestrophin-1 expression, localisation and function in a polarised epithelial cell model.
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DOI：
10.1242/dmm.024216
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发表时间：
2016-11-01
影响因子：
4.3
通讯作者：
Manson FD
中科院分区：
文献类型：
--
作者：
Uggenti C;Briant K;Streit AK;Thomson S;Koay YH;Baines RA;Swanton E;Manson FD
常染色体隐性雌激素样蛋白病（ARB）是一种由雌激素样蛋白-1突变引起的视网膜病，被认为是视网膜色素上皮（RPE）基底外侧表面的Ca 2+门控Cl−通道。使用稳定转染的极化上皮细胞模型，我们表明，四个ARB突变体雌激素-1蛋白被错误定位，并进行蛋白酶体降解。与野生型bestrophin-1相反，四种突变蛋白中的每一种都不能在瞬时转染的细胞中传导Cl−离子，这是通过全细胞膜片钳测定的。我们证明，两种临床批准的药物，硼替佐米和4-苯基丁酸（4PBA）的组合，成功地恢复了所有四种ARB突变体雌激素-1蛋白的表达和定位。重要的是，通过单独用4PBA处理细胞，每种突变型雌激素样蛋白-1的Cl−传导功能都完全恢复到野生型雌激素样蛋白-1的功能。使用4PBA实现的功能拯救是重要的，因为它表明这种已经被批准用于婴儿和成人长期使用的药物可能代表了治疗ARB和BEST 1错义突变引起的其他雌激素病的有希望的疗法。总结：化学伴侣4PBA完全恢复与遗传性视网膜营养不良、常染色体隐性雌激素样蛋白病相关的突变型雌激素样蛋白-1蛋白的Cl−传导活性。
Autosomal recessive bestrophinopathy (ARB) is a retinopathy caused by mutations in the bestrophin-1 protein, which is thought to function as a Ca2+-gated Cl− channel in the basolateral surface of the retinal pigment epithelium (RPE). Using a stably transfected polarised epithelial cell model, we show that four ARB mutant bestrophin-1 proteins were mislocalised and subjected to proteasomal degradation. In contrast to the wild-type bestrophin-1, each of the four mutant proteins also failed to conduct Cl− ions in transiently transfected cells as determined by whole-cell patch clamp. We demonstrate that a combination of two clinically approved drugs, bortezomib and 4-phenylbutyrate (4PBA), successfully restored the expression and localisation of all four ARB mutant bestrophin-1 proteins. Importantly, the Cl− conductance function of each of the mutant bestrophin-1 proteins was fully restored to that of wild-type bestrophin-1 by treatment of cells with 4PBA alone. The functional rescue achieved with 4PBA is significant because it suggests that this drug, which is already approved for long-term use in infants and adults, might represent a promising therapy for the treatment of ARB and other bestrophinopathies resulting from missense mutations in BEST1. Summary: Chemical chaperone 4PBA fully restores Cl− conductance activity for mutant bestrophin-1 proteins associated with inherited retinal dystrophy, autosomal recessive bestrophinopathy.
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DOI：
10.1111/j.1600-0854.2010.01155.x
发表时间：
2011-04
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
Castorino JJ;Deborde S;Deora A;Schreiner R;Gallagher-Colombo SM;Rodriguez-Boulan E;Philp NJ
通讯作者：
Philp NJ
影响因子：
2.8
作者：
Field-Smith A;Morgan GJ;Davies FE
通讯作者：
Davies FE
影响因子：
5.6
作者：
Doumanov JA;Zeitz C;Dominguez Gimenez P;Audo I;Krishna A;Alfano G;Diaz ML;Moskova-Doumanova V;Lancelot ME;Sahel JA;Nandrot EF;Bhattacharya SS
通讯作者：
Bhattacharya SS
影响因子：
5.1
作者：
Batshaw, ML;MacArthur, RB;Tuchman, M
通讯作者：
Tuchman, M
影响因子：
13.7
作者：
Boon, Camiel J. F.;van den Born, L. Ingeborgh;van Schooneveld, Mary J.
通讯作者：
van Schooneveld, Mary J.