Title
In Situ Cyclization of Proteins (INCYPRO): Cross-Link Derivatization Modulates Protein Stability
Author
Saskia Neubacher, Jordy M. Saya, Alessia Amore, Tom N. Grossmann
Year
2019
Journal
The Journal of Organic Chemistry
Abstract
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.
Full Article
Instrument
J-1500
Keywords
Circular dichroism, Secondary structure, Ligand binding, Pharmaceutical, Biochemistry