GwAAP: A genome-wide amino acid coding-decoding quantitative proteomics system

Abstract

Summary: Mass spectrometry-based proteomic technology has greatly improved and has been widely applied in various biological science fields. However, proteome-wide accurate quantification of proteins in signaling pathways remains challenging. Here, we report a genome-wide amino acid coding-decoding quantitative proteomic (GwAAP) system to facilitate precise proteome quantification. For each protein, a unique code peptide was assigned and incorporated into the N-terminus of the targeted protein and used for identification and quantification. As a proof of principle, we systematically tagged 40 yeast proteins with codes and employed mass spectrometry to decode. We successfully recovered all 40 code peptides with a large and consistent quantitative dynamic range (CV slope 0.8). We further verified the alteration of the glucose and galactose metabolism pathways in yeast under different carbon source conditions. The GwAAP system could potentially provide a strategy to achieve absolute quantification of the entire yeast proteome without bias.

Keywords

• Biochemistry methods
• Biological sciences research methodologies
• Proteomics