In Situ Cyclization of Proteins (INCYPRO): Cross-Link Derivatization Modulates Protein Stability

Saskia Neubacher*, Jordy M. Saya, Alessia Amore, Tom N. Grossmann

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

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Protein macrocyclization represents a very efficient strategy to increase the stability of protein tertiary structures. Here, we describe a panel of novel C3-symmetric tris-electrophilic agents and their use for the cyclization of proteins. These electrophiles are reacted with a protein domain harboring three solvent-exposed cysteine residues, resulting in the in situ cyclization of the protein (INCYPRO). We observe a clear dependency of cross-linking rates on the electrophilicity. All nine obtained cross-linked protein versions show considerably increased thermal stability (up to 29 °C increased melting temperature) when compared to that of the linear precursor. Most interestingly, the degree of stabilization correlates with the hydrophilicity of the cross-link. These results will support the development of novel cross-linked proteins and enable a more rational design process.

Original languageEnglish
Pages (from-to)1476-1483
Number of pages8
JournalJournal of Organic Chemistry
Issue number3
Early online date2 Dec 2019
Publication statusPublished - 7 Feb 2020


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