Cells | |
Quantification of High-Molecular Weight Protein Platforms by AQUA Mass Spectrometry as Exemplified for the CD95 Death-Inducing Signaling Complex (DISC) | |
Uwe Warnken2  Kolja Schleich1  Martina Schnölzer2  | |
[1] Division of Immunogenetics, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany;Functional Proteome Analysis, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany | |
关键词: CD95; DISC; apoptosis; DED; protein quantification; mass spectrometry; AQUA technique; protein platforms; | |
DOI : 10.3390/cells2030476 | |
来源: mdpi | |
【 摘 要 】
Contemporary quantitative mass spectrometry provides fascinating opportunities in defining the stoichiometry of high-molecular weight complexes or multiprotein platforms. The composition stoichiometry of multiprotein platforms is a key to understand the regulation of complex signaling pathways and provides a basis for constructing models in systems biology. Here we present an improved AQUA technique workflow that we adapted for the quantitative mass spectrometry analysis of the stoichiometry of the CD95 (Fas/APO-1) death inducing signaling complex (DISC). The DISC is a high-molecular weight platform essential for the initiation of CD95-mediated apoptotic and non-apoptotic responses. For protein quantification, CD95 DISCs were immunoprecipitated and proteins in the immunoprecipitations were separated by one-dimensional gel electrophoresis, followed by protein quantification using the AQUA technique. We will discuss in detail AQUA analysis of the CD95 DISC focusing on the key issues of this methodology,
【 授权许可】
CC BY
© 2013 by the authors; licensee MDPI, Basel, Switzerland.
【 预 览 】
Files | Size | Format | View |
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RO202003190035466ZK.pdf | 2075KB | download |