Monitoring Disassembly and Cargo Release of Phase-Separated Peptide Coacervates with Native Mass Spectrometry

Cerrato, Carmine P. P.Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Biomedicum, S-17165 Solna, Sweden (författare)

Engineering liquid-liquid phase separation (LLPS)of proteinsand peptides holds great promise for the development of therapeuticcarriers with intracellular delivery capability but requires accuratedetermination of their assembly properties in vitro, usually with fluorescently labeled cargo. Here, we use mass spectrometry(MS) to investigate redox-sensitive coacervate microdroplets (thedense phase formed during LLPS) assembled from a short His- and Tyr-richpeptide. We can monitor the enrichment of a reduced peptide in dilutephase as the microdroplets dissolve triggered by their redox-sensitiveside chain, thus providing a quantitative readout for disassembly.Furthermore, MS can detect the release of a short peptide from coacervatesunder reducing conditions. In summary, with MS, we can monitor thedisassembly and cargo release of engineered coacervates used as therapeuticcarriers without the need for additional labels.

Leppert, AxelKarolinska Inst, Dept Microbiol Tumor & Cell Biol, Biomedicum, S-17165 Solna, Sweden (författare)
Sun, YueNanyang Technol Univ NTU, Ctr Sustainable Mat SusMat, Sch Mat Sci & Engn, Biol & Biomimet Mat Lab BBML, Singapore 637553, Singapore (författare)
Lane, David P. P.Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Biomedicum, S-17165 Solna, Sweden (författare)
Arsenian-Henriksson, MarieKarolinska Institutet (författare)
Miserez, AliNanyang Technol Univ NTU, Ctr Sustainable Mat SusMat, Sch Mat Sci & Engn, Biol & Biomimet Mat Lab BBML, Singapore 637553, Singapore.;Nanyang Technol Univ NTU, Sch Biol Sci, Singapore 637553, Singapore (författare)
Landreh, MichaelKarolinska Inst, Dept Microbiol Tumor & Cell Biol, Biomedicum, S-17165 Solna, Sweden(Swepub:uu)micla833 (författare)
Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Biomedicum, S-17165 Solna, SwedenNanyang Technol Univ NTU, Ctr Sustainable Mat SusMat, Sch Mat Sci & Engn, Biol & Biomimet Mat Lab BBML, Singapore 637553, Singapore (creator_code:org_t)

Ingår i:Analytical Chemistry: American Chemical Society (ACS)95:29, s. 10869-108720003-27001520-6882

Av författaren/redakt...: Cerrato, Carmine ...; Leppert, Axel; Sun, Yue; Lane, David P. P ...; Arsenian-Henriks ...; Miserez, Ali; visa fler...; Landreh, Michael; visa färre...

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