用于溶酶体分离的超顺磁纳米粒子，以鉴定溶酶体蛋白和脂质组成的空间变化。

Superparamagnetic Nanoparticles for Lysosome Isolation to Identify Spatial Alterations in Lysosomal Protein and Lipid Composition.

机构信息

Laboratory of Neurobiology, VIB-Center for Brain & Disease Research, Leuven, Belgium.

Department of Neurosciences, KU Leuven, Leuven, Belgium.

出版信息

STAR Protoc. 2020 Oct 9;1(3):100122. doi: 10.1016/j.xpro.2020.100122. eCollection 2020 Dec 18.

DOI:10.1016/j.xpro.2020.100122

PMID:33377016

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7756972/

Abstract

Lysosomes are dynamic organelles that serve as regulatory hubs in cellular homeostasis. Changes in lysosome morphology, composition, and turnover are typically linked to disease. These characteristics make enrichment protocols based on biophysical parameters challenging. However, organelle enrichment methods are essential to facilitate their biomolecular analysis. We describe the synthesis and use of superparamagnetic iron oxide nanoparticles (SPIONs) for high-yield purification of lysosomes compatible with "omics" analysis. NANOLYSE (Nanoparticles for Lysosome Isolation) provides a reliable strategy in fingerprinting the biomolecular composition of lysosomes. For complete details on the use and execution of this protocol, please refer to Tharkeshwar et al. (2017).

摘要

溶酶体是具有动态性的细胞器，在细胞内稳态中充当调节枢纽。溶酶体形态、组成和周转率的变化通常与疾病有关。这些特征使得基于生物物理参数的溶酶体富集方案具有挑战性。然而，细胞器的富集方法对于促进它们的生物分子分析是必不可少的。我们描述了超顺磁性氧化铁纳米粒子（SPIONs）的合成和使用，用于高效纯化适合“组学”分析的溶酶体。NANOLYSE（用于溶酶体分离的纳米颗粒）为指纹分析溶酶体的生物分子组成提供了一种可靠的策略。有关此方案使用和执行的完整详细信息，请参阅 Tharkeshwar 等人。（2017 年）。