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Nat Methods. 2011 Jul 10;8(8):691-6. doi: 10.1038/nmeth.1649.

Protein standard absolute quantification (PSAQ) method for the measurement of cellular ubiquitin pools.

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1
Department of Biology, Stanford University, Stanford, California, USA.

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.

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PMID:
21743460
PMCID:
PMC3196335
DOI:
10.1038/nmeth.1649
[Indexed for MEDLINE]
Free PMC Article

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