Journal article

Publication year: 2012

Pages: 512-526

Journal: Molecular & Cellular Proteomics

Volume number: 11

Issue number: 8

ISSN: 1535-9476

eISSN: 1535-9484

DOI Link: https://doi.org/10.1074/mcp.M111.014134

Publisher: Elsevier

Abstract: 
Liquid chromatography coupled to tandem mass spectrometry in combination with stable-isotope labeling is an established and widely spread method to measure gene expression on the protein level. However, it is often not considered that two opposing processes are responsible for the amount of a protein in a cell-the synthesis as well as the degradation. With this work, we provide an integrative, high-throughput method-from the experimental setup to the bioinformatics analysis-to measure synthesis and degradation rates of an organism's proteome. Applicability of the approach is demonstrated with an investigation of heat shock response, a well-understood regulatory mechanism in bacteria, on the biotechnologically relevant Corynebacterium glutamicum. Utilizing a multilabeling approach using both heavy stable nitrogen as well as carbon isotopes cells are metabolically labeled in a pulse-chase experiment to trace the labels' incorporation in newly synthesized proteins and its loss during protein degradation. Our work aims not only at the calculation of protein turnover rates but also at their statistical evaluation, including variance and hierarchical cluster analysis using the rich internet application QuPE.

Harvard Citation style: Trötschel, C., Albaum, S., Wolff, D., Schröder, S., Goesmann, A., Nattkemper, T., et al. (2012) Protein Turnover Quantification in a Multilabeling Approach: From Data Calculation to Evaluation, Molecular & Cellular Proteomics, 11(8), pp. 512-526. https://doi.org/10.1074/mcp.M111.014134

APA Citation style: Trötschel, C., Albaum, S., Wolff, D., Schröder, S., Goesmann, A., Nattkemper, T., & Poetsch, A. (2012). Protein Turnover Quantification in a Multilabeling Approach: From Data Calculation to Evaluation. Molecular & Cellular Proteomics. 11(8), 512-526. https://doi.org/10.1074/mcp.M111.014134