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High-Throughput Proteomics Enabled by a Photocleavable Surfactant.

Angewandte Chemie (International ed. in English) (2020-02-26)
Kyle A Brown, Trisha Tucholski, Christian Eken, Samantha Knott, Yanlong Zhu, Song Jin, Ying Ge
RESUMEN

Mass spectrometry (MS)-based proteomics provides unprecedented opportunities for understanding the structure and function of proteins in complex biological systems; however, protein solubility and sample preparation before MS remain a bottleneck preventing high-throughput proteomics. Herein, we report a high-throughput bottom-up proteomic method enabled by a newly developed MS-compatible photocleavable surfactant, 4-hexylphenylazosulfonate (Azo) that facilitates robust protein extraction, rapid enzymatic digestion (30 min compared to overnight), and subsequent MS-analysis following UV degradation. Moreover, we developed an Azo-aided bottom-up method for analysis of integral membrane proteins, which are key drug targets and are generally underrepresented in global proteomic studies. Furthermore, we demonstrated the ability of Azo to serve as an "all-in-one" MS-compatible surfactant for both top-down and bottom-up proteomics, with streamlined workflows for high-throughput proteomics amenable to clinical applications.

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Sigma-Aldrich
Sodium 2-(4-hexylphenyl)diazene-1-sulfonate, (0.5% solution in 25mM aqueous ammonium bicarbonate, pH 7.0), ≥95%