TY - JOUR
T1 - Monitoring Disassembly and Cargo Release of Phase-Separated Peptide Coacervates with Native Mass Spectrometry
AU - Cerrato, Carmine P.
AU - Leppert, Axel
AU - Sun, Yue
AU - Lane, David P.
AU - Arsenian-Henriksson, Marie
AU - Miserez, Ali
AU - Landreh, Michael
N1 - Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society.
PY - 2023/7/25
Y1 - 2023/7/25
N2 - 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.
AB - 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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U2 - 10.1021/acs.analchem.3c02384
DO - 10.1021/acs.analchem.3c02384
M3 - Article
AN - SCOPUS:85165692927
SN - 0003-2700
VL - 95
SP - 10869
EP - 10872
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 29
ER -