Monitoring disassembly and cargo release of phase-separated peptide coacervates with native mass spectrometry

Monitoring disassembly and cargo release of phase-separated peptide coacervates with native mass spectrometry

Engineering liquid-liquid phase separation (LLPS) of proteins and peptides holds great promise for the development of therapeutic carriers with intracellular delivery capability but requires accurate determination of their assembly properties in vitro, usually with fluorescently labeled cargo. Here,...

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Bibliographic Details
Main Authors: Cerrato, Carmine P., Leppert, Axel, Sun, Yue, Lane, David P., Arsenian-Henriksson, Marie, Miserez, Ali, Landreh, Michael
Other Authors: School of Materials Science and Engineering
Format: Article
Language:English
Published: 2023
Subjects:
Engineering::Materials
Native Mass Spectrometry
Peptide Coacervates
Online Access:https://hdl.handle.net/10356/171059
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Institution: Nanyang Technological University
Language: English
Description
Summary:Engineering liquid-liquid phase separation (LLPS) of proteins and peptides holds great promise for the development of therapeutic carriers with intracellular delivery capability but requires accurate determination of their assembly properties in vitro, usually with fluorescently labeled cargo. Here, we use mass spectrometry (MS) to investigate redox-sensitive coacervate microdroplets (the dense phase formed during LLPS) assembled from a short His- and Tyr-rich peptide. We can monitor the enrichment of a reduced peptide in dilute phase as the microdroplets dissolve triggered by their redox-sensitive side chain, thus providing a quantitative readout for disassembly. Furthermore, MS can detect the release of a short peptide from coacervates under reducing conditions. In summary, with MS, we can monitor the disassembly and cargo release of engineered coacervates used as therapeutic carriers without the need for additional labels.

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