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


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.

SUBMITTER: Kaiser SE 

PROVIDER: S-EPMC3196335 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature

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

Kaiser Stephen E SE   Riley Brigit E BE   Shaler Thomas A TA   Trevino R Sean RS   Becker Christopher H CH   Schulman Howard H   Kopito Ron R RR  

Nature methods 20110710 8


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 cellu  ...[more]

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