Thickness determination of hydroperoxidized lipid bilayers from medium-resolution cryo-TEM images.

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology
Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-04-23 DOI:10.1016/bs.mie.2024.02.008
Eulalie Lafarge, Carlos M Marques, Marc Schmutz, Pierre Muller, André P Schroder
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引用次数: 0

Abstract

As the primary products of lipid oxidation, lipid hydroperoxides constitute an important class of lipids generated by aerobic metabolism. However, despite several years of effort, the structure of the hydroperoxidized bilayer has not yet been observed under electron microscopy. Here we use a 200 kV Cryo-TEM to image small unilamellar vesicles (SUVs) made (i) of pure POPC or SOPC, (ii) of their pure hydroperoxidized form, and (iii) of their equimolar mixtures. We show that the challenges posed by the determination of the thickness of the hydroperoxidized bilayers under these observation conditions can be addressed by an image analysis method that we developed and describe here.

从中等分辨率的低温 TEM 图像中确定氢过氧化脂质双分子层的厚度。
作为脂质氧化的主要产物,脂质氢过氧化物是有氧代谢产生的一类重要脂质。然而,尽管经过数年的努力,氢过氧化物双分子层的结构仍未在电子显微镜下观察到。在此,我们使用 200 kV Cryo-TEM 对小型单酰胺囊泡 (SUV) 进行成像:(i) 纯 POPC 或 SOPC,(ii) 纯氢过氧化物,(iii) 它们的等摩尔混合物。我们的研究表明,在这些观察条件下测定氢过氧化双分子层厚度所面临的挑战可以通过我们开发并在此介绍的一种图像分析方法来解决。
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来源期刊
Methods in enzymology
Methods in enzymology 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
308
审稿时长
3-6 weeks
期刊介绍: The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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