Protein standard absolute quantification (PSAQ) method for the measurement of cellular ubiquitin pools |
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Authors: | Kaiser Stephen E Riley Brigit E Shaler Thomas A Trevino R Sean Becker Christopher H Schulman Howard Kopito Ron R |
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Institution: | Department of Biology, Stanford University, Stanford, California, USA. |
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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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