Codetermination of Sphingomyelin and Cholesterol in Cellular Plasma Membrane in Sphingomyelin-Depletion-Induced Cholesterol Efflux.,Analytical Chemistry

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Codetermination of Sphingomyelin and Cholesterol in Cellular Plasma Membrane in Sphingomyelin-Depletion-Induced Cholesterol Efflux.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-12-14 00:00:00 , DOI: 10.1021/acs.analchem.8b04593
Shuohan Huang ₁ , Kang Liu ₁ , Dechen Jiang ₂ , Danjun Fang ₁

Affiliation

Quantification of multiple lipids with different contents in plasma membrane in single cells is significant, but challenging, for investigating lipid interactions and the role of dominant protein transporters. In this paper, comonitoring the alteration of low-content sphingomyelin (SM) and high-content cholesterol in plasma membrane of one living cell is realized by use of luminol electrochemiluminescence (ECL) for the first time. Concentrations of SM as low as 0.5 μM are detected, which permits the measurement of low-content membrane SM in single cells. More membrane cholesterol is observed in individual cells after depletion of membrane SM, providing direct evidence about SM-depletion-induced cholesterol efflux. The upregulation of ATP-binding cassette transporters A1 (ABCA1) and G1 (ABCG1) in SM-depleted cells induces a further increase in membrane cholesterol. These results imply that higher expression of ABCA1/G1 promotes cholesterol trafficking, which offers additional information to solve the debate about ABC transporters in cholesterol efflux. Moreover, the established approach offers a special strategy to investigate the correlation of membrane lipids and the role of transporters in cholesterol trafficking.

中文翻译：

鞘磷脂消耗诱导的胆固醇外流的细胞质膜中鞘磷脂和胆固醇的共解。

对于研究脂质相互作用和主要蛋白转运蛋白的作用，单细胞质膜中不同含量的多种脂质的定量分析是重要的，但具有挑战性。本文首次通过使用鲁米诺电化学发光（ECL）来实现对一个活细胞质膜中低含量的鞘磷脂（SM）和高含量胆固醇的改变的监测。检测到的SM浓度低至0.5μM，从而可以测量单个细胞中的低含量膜SM。膜SM耗尽后，在单个细胞中观察到更多的膜胆固醇，这提供了有关SM耗尽诱导的胆固醇外流的直接证据。SM缺失的细胞中ATP结合盒转运蛋白A1（ABCA1）和G1（ABCG1）的上调诱导膜胆固醇进一步增加。这些结果表明，ABCA1 / G1的高表达促进胆固醇的运输，这为解决有关ABC转运蛋白在胆固醇外流中的争论提供了更多信息。此外，已建立的方法提供了一种特殊的策略来研究膜脂的相关性以及转运蛋白在胆固醇运输中的作用。

更新日期：2018-12-14

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