用于测量细胞泛素池的蛋白质标准绝对定量 (PSAQ) 方法。
Protein standard absolute quantification (PSAQ) method for the measurement of cellular ubiquitin pools.
机构信息
Department of Biology, Stanford University, Stanford, California, USA.
出版信息
Nat Methods. 2011 Jul 10;8(8):691-6. doi: 10.1038/nmeth.1649.
Abstract
The protein ubiquitin is an important post-translational modifier that regulates a wide variety of biological processes. In cells, ubiquitin is apportioned among distinct pools, which include a variety of free and conjugated species. Although maintenance of a dynamic and complex equilibrium among ubiquitin pools is crucial for cell survival, the tools necessary to quantify each cellular ubiquitin pool have been limited. We have developed a quantitative mass spectrometry approach to measure cellular concentrations of ubiquitin species using isotope-labeled protein standards and applied it to characterize ubiquitin pools in cells and tissues. Our method is convenient, adaptable and should be a valuable tool to facilitate our understanding of this important signaling molecule.
摘要
蛋白质泛素是一种重要的翻译后修饰物，可调节多种生物过程。在细胞中，泛素被分配到不同的池中，包括多种游离和缀合的物质。尽管保持泛素池之间的动态和复杂平衡对于细胞存活至关重要，但量化每个细胞泛素池的工具一直受到限制。我们开发了一种定量质谱方法，使用同位素标记的蛋白质标准品来测量细胞中泛素物质的浓度，并将其应用于细胞和组织中泛素池的特征描述。我们的方法简便、适应性强，应该是促进我们理解这种重要信号分子的有价值的工具。
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