Program in Integrative Nutrition and Complex Diseases, Texas A&M University, College Station, TX, USA.
Department of Nutrition, Texas A&M University, College Station, TX, USA.
Methods Mol Biol. 2021;2262:251-258. doi: 10.1007/978-1-0716-1190-6_14.
Phospholipid fatty acid (FA) composition influences the biophysical properties of the plasma membrane and plays an important role in cellular signaling. Our previous work has demonstrated that plasma membrane fatty acid composition is an important determinant of oncogenic Ras signaling and that dietary (exogenous) modulation of membrane composition may underlie the chemoprotective benefits of long chain n-3 polyunsaturated fatty acids (PUFA). In this chapter, we describe in vitro methods to modulate membrane phospholipid fatty acid composition of cultured cells using fatty acids complexed to bovine serum albumin (BSA). Furthermore, we describe a method to quantify the biophysical properties of plasma membranes in live cells using Di-4-ANEPPDHQ (Di4) and image-based flow cytometry.
磷脂脂肪酸（FA）组成影响质膜的生物物理特性，并在细胞信号转导中发挥重要作用。我们之前的工作表明，质膜脂肪酸组成是致癌 Ras 信号的重要决定因素，并且膜组成的饮食（外源性）调节可能是长链 n-3 多不饱和脂肪酸（PUFA）的化学保护益处的基础。在本章中，我们描述了使用与牛血清白蛋白（BSA）复合的脂肪酸来调节培养细胞中膜磷脂脂肪酸组成的体外方法。此外，我们描述了一种使用 Di-4-ANEPPDHQ（Di4）和基于图像的流式细胞术定量活细胞中质膜生物物理特性的方法。
