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Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
Published on: November 28, 2017
Electrostatically self-assembled azides on zinc sulfide nanoparticles as multifunctional nanoprobes for peptide and
Hui-Fen Wu1, Suresh Kumar Kailasa, Lokesh Shastri
1Department of Chemistry, National Sun Yat-Sen University, Kaohsiung, Taiwan. hwu@faculty.nsysu.edu.tw
Abstract:
A simple method to synthesize electrostatically self-assembled azides on zinc sulfide nanoparticles (ZnS-N(3) NPs) was described and then it was further applied as a multifunctional nanoprobe such as enriching, desalting, accelerating and separation-/washing free nanoprobes for rapid analysis of peptides and proteins and microwave assisted tryptic digested proteins in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The ZnS-N(3) NPs were characterized by UV-vis, FT-IR, SEM and TEM spectroscopy. The ZnS-N(3) NPs can effectively enrich signal intensities for 2-10 times for various peptides and proteins including HW6, insulin, ubiquitin, cytochrome c, lysozyme, myoglobin and bovine serum albumin (BSA) in MALDI-TOF MS. Furthermore, we also demonstrated that the ZnS-N(3) NPs can serve as accelerating probes for microwave assisted tryptic digestion of proteins in MALDI-TOF MS. The applicability of the present method on complex sample analysis such as milk proteins from cow milk and ubiquitin and ubiquitin like proteins from oyster mushroom were also demonstrated.
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