Who's in, who's out? Re-evaluation of lipid raft residents.
Who's in, who's out? Re-evaluation of lipid raft residents.
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DOI：
10.1111/jnc.15446
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发表时间：
2021-08
影响因子：
4.7
通讯作者：
Heffer M
中科院分区：
文献类型：
--
作者：
Mlinac-Jerkovic K;Ilic K;Zjalić M;Mandić D;Debeljak Ž;Balog M;Damjanović V;Maček Hrvat N;Habek N;Kalanj-Bognar S;Schnaar RL;Heffer M
脂筏，富含（糖）鞘脂，胆固醇和选择蛋白质的膜微区，作为细胞信号体。已使用各种方法从本体（非筏）膜中分离脂筏，但最常见的是使用非离子去污剂Triton X-100进行分离。然而，Triton X-100是一种脂筏破坏剂。组织学证据证实了Triton X-100对筏的破坏，但显著显示了用相关聚环氧乙烷洗涤剂Brij O20处理的筏稳定性。我们报告了使用Brij O20从小鼠脑中分离抗洗涤剂膜，并使用其确定主要哺乳动物脑神经节苷脂GM 1，GD 1a，GD 1b和GT 1b的分布。使用Brij O20与Triton X-100发现了神经节苷脂的不同分布-经典上用作筏标记物。免疫组织化学和成像质谱证实了这一结果。使用Brij O20导致神经节苷脂的独特膜分布，其并非全部与脂筏相关，而是取决于神经节苷脂结构。这是第一次报告的一个重要比例的神经节苷脂外筏域。我们还确定了与神经可塑性功能相关的蛋白质的分布，并已知受神经节苷脂环境，谷氨酸受体亚基2，淀粉样前体蛋白和neuroplastin的影响，并报告了这些蛋白质在小鼠脑组织中的脂筏种群。这项工作将使更准确的脂筏分析鞘糖脂和膜蛋白的组成，并导致生物膜内的脂质-蛋白质相互作用的分辨率提高。使用数据驱动的分区策略，我们报告说，一个显着比例的神经节苷脂，长期以来被认为是脂筏居民，居住在筏域之外。免疫组织化学和成像质谱显示，与广泛使用的Triton X-100（诱导神经节苷脂重新分布）相比，Brij O20去污剂处理后神经节苷脂的分布基本保持不变。Brij O20脑提取物的脂筏分析表明，神经节苷脂并不都是脂筏，这取决于神经节苷脂的结构。同样，不同的神经可塑性蛋白筏分布揭示。这项工作使更准确的脂筏分析的生物膜中的脂质-蛋白质相互作用的决议。
Lipid rafts, membrane microdomains enriched with (glyco)sphingolipids, cholesterol, and select proteins, act as cellular signalosomes. Various methods have been used to separate lipid rafts from bulk (non‐raft) membranes, but most often, non‐ionic detergent Triton X‐100 has been used in their isolation. However, Triton X‐100 is a reported disruptor of lipid rafts. Histological evidence confirmed raft disruption by Triton X‐100, but remarkably revealed raft stability to treatment with a related polyethylene oxide detergent, Brij O20. We report isolation of detergent‐resistant membranes from mouse brain using Brij O20 and its use to determine the distribution of major mammalian brain gangliosides, GM1, GD1a, GD1b and GT1b. A different distribution of gangliosides—classically used as a raft marker—was discovered using Brij O20 versus Triton X‐100. Immunohistochemistry and imaging mass spectrometry confirm the results. Use of Brij O20 results in a distinctive membrane distribution of gangliosides that is not all lipid raft associated, but depends on the ganglioside structure. This is the first report of a significant proportion of gangliosides outside raft domains. We also determined the distribution of proteins functionally related to neuroplasticity and known to be affected by ganglioside environment, glutamate receptor subunit 2, amyloid precursor protein and neuroplastin and report the lipid raft populations of these proteins in mouse brain tissue. This work will enable more accurate lipid raft analysis with respect to glycosphingolipid and membrane protein composition and lead to improved resolution of lipid–protein interactions within biological membranes. Using a data‐driven partitioning strategy, we report that a significant proportion of gangliosides, long believed to be lipid raft resident, reside outside raft domains. Immunohistochemistry and imaging mass spectrometry revealed that the distribution of gangliosides remains largely unchanged by treatment with Brij O20 detergent compared to the widely used Triton X‐100, which induces ganglioside redistribution. Lipid raft analyses of Brij O20 brain extracts revealed that gangliosides are not all lipid raft depending on the ganglioside structures. Likewise, different neuroplasticity protein raft distributions were revealed. This work enables more accurate lipid raft analyses for resolution of lipid–protein interactions in biological membranes.
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影响因子：
4
作者：
Hicks DA;Nalivaeva NN;Turner AJ
通讯作者：
Turner AJ
影响因子：
5.5
作者：
Ilic, Katarina;Auer, Benedikt;Herrera-Molina, Rodrigo
通讯作者：
Herrera-Molina, Rodrigo
影响因子：
2.9
作者：
Eckert, GP;Igbavboa, U;Wood, WG
通讯作者：
Wood, WG
影响因子：
4.8
作者：
Herrera-Molina, Rodrigo;Sarto-Jackson, Isabella;Montag, Dirk
通讯作者：
Montag, Dirk
影响因子：
4.6
作者：
Herrera-Molina R;Mlinac-Jerkovic K;Ilic K;Stöber F;Vemula SK;Sandoval M;Milosevic NJ;Simic G;Smalla KH;Goldschmidt J;Bognar SK;Montag D
通讯作者：
Montag D