High‐Throughput Proteomics Enabled by a Photocleavable Surfactant. Issue 22 (24th March 2020)

• Record Type: Journal Article
• Authors: Brown, Kyle A.; Tucholski, Trisha; Eken, Christian; Knott, Samantha; Zhu, Yanlong; Jin, Song; Ge, Ying
• Is Part Of: Angewandte Chemie international edition. Volume 59:Issue 22(2020)
• Page Start: 8406
• Page End: 8410
• Publication Date: 2020-03-24
• Subjects: high-throughput proteomics -- mass spectrometry -- membrane proteins -- photocleavable surfactants; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201915374
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: 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. Abstract : Photocleavable surfactant : The unique ability of 4‐hexylphenylazosulfonate (Azo) to serve as an "all‐in‐one" MS‐compatible surfactant for both high‐throughput bottom‐up and top‐down proteomics is demonstrated.