Proteome Dynamics with Heavy Water - Instrumentations, Data Analysis, and Biological Applications
The quantitative assessment of the synthesis of individual proteins has been greatly hindered by the lack of a high-throughput nonradioactive method. We recently developed a method that we call "proteome dynamics" and software that enables high-throughput kinetic analyses of peptides on a...
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| Main Authors: | , , , , |
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| Format: | Ebooks |
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IntechOpen,
2015-11-11.
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| Online Access: | Get Online |
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| Summary: | The quantitative assessment of the synthesis of individual proteins has been greatly hindered by the lack of a high-throughput nonradioactive method. We recently developed a method that we call "proteome dynamics" and software that enables high-throughput kinetic analyses of peptides on a proteome-wide scale. Previous studies established that oral administration of heavy water (2H2O or deuterium oxide, D2O) is safe and well tolerated in humans. Briefly, a loading dose of 2H2O, a nonradioactive isotope, is administered in drinking water. 2H2O rapidly labels body water and transfers 2H from 2H2O to 2H-labeled amino acids, which incorporates into proteins dependent upon the rate of synthesis of the specific protein. Proteins are analyzed by high-resolution mass spectrometry and protein synthesis is calculated using specialized software. We have established the effectiveness of this method for plasma and mitochondrial proteins. We demonstrated that fasting has a differential effect on the synthesis rates of proteins. We also applied this method to assess the effect of heart failure on the stability of mitochondrial proteins. In this review, we describe the study design, instrumentation, data analysis, and biological application of heavy water-based proteome turnover studies. We summarize this chapter with the challenges in the field and future directions. |
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| Item Description: | https://mts.intechopen.com/articles/show/title/proteome-dynamics-with-heavy-water-instrumentations-data-analysis-and-biological-applications |