Su Xue Bessie, Mudhaffer Amina, Saiardi Adolfo
Laboratory for Molecular Cell Biology, University College London, London, UK.
Methods Mol Biol. 2025;2972:19-27. doi: 10.1007/978-1-0716-4799-8_2.
Inositol polyphosphates are ubiquitously present in eukaryotic cells and play important roles in diverse cellular processes such as protein folding, signal transduction, and phosphate homeostasis. Although these negatively charged small metabolites are easily stripped from metals/proteins and extracted under strong acidic conditions, their low intracellular concentrations in some biological settings command a further enrichment step to facilitate downstream analyses. Here, we describe the use of titanium dioxide beads to enrich for inositol polyphosphate extracted from cultured mammalian cells. This protocol not only concentrates the diluted inositol phosphates but also removes salts and proteins that would otherwise interfere with postpurification analyses.
肌醇多磷酸广泛存在于真核细胞中，在多种细胞过程中发挥重要作用，如蛋白质折叠、信号转导和磷酸盐稳态。尽管这些带负电荷的小代谢物很容易从金属/蛋白质上剥离，并在强酸性条件下提取，但在某些生物学环境中它们的细胞内浓度较低，需要进一步富集步骤以利于下游分析。在此，我们描述了使用二氧化钛珠富集从培养的哺乳动物细胞中提取的肌醇多磷酸。该方案不仅能浓缩稀释后的肌醇磷酸，还能去除否则会干扰纯化后分析的盐和蛋白质。
