Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

Author(s):Show less -

Bernardim, Barbara ; Cal, Pedro M. S. D. ; Matos, Maria J. ; Oliveira, Bruno L. ; Martinez-Saez, Nuria ; Albuquerque, Ines S. ; Perkins, Elizabeth ; Corzana, Francisco ; Burtoloso, Antonio C. B. ; Jimenez-Oses, Gonzalo ; Bernardes, Goncalo J. L.

• Document type: Journal article
• Source: NATURE COMMUNICATIONS; v. 7, OCT 26 2016.
• Web of Science Citations: 31

Abstract

Maleimides remain the reagents of choice for the preparation of therapeutic and imaging protein conjugates despite the known instability of the resulting products that undergo thiol-exchange reactions in vivo. Here we present the rational design of carbonylacrylic reagents for chemoselective cysteine bioconjugation. These reagents undergo rapid thiol Michael-addition under biocompatible conditions in stoichiometric amounts. When using carbonylacrylic reagents equipped with PEG or fluorophore moieties, this method enables access to protein and antibody conjugates precisely modified at pre-determined sites. Importantly, the conjugates formed are resistant to degradation in plasma and are biologically functional, as demonstrated by the selective imaging and detection of apoptotic and HER2+ cells, respectively. The straightforward preparation, stoichiometric use and exquisite cysteine selectivity of the carbonylacrylic reagents combined with the stability of the products and the availability of biologically relevant cysteine-tagged proteins make this method suitable for the routine preparation of chemically defined conjugates for in vivo applications. (AU)

• FAPESP's process: 15/07509-1 - Site-specific modification of proteins via aqueous Horner-Wadsworth-Emmons reaction followed by Wolff rearrangement
• Grantee: Barbara Bernardim de Souza
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 13/25504-1 - Development of new methodologies for the asymmetric a-functionalization of carbonyl compounds using chiral catalysts
• Grantee: Antonio Carlos Bender Burtoloso
• Support Opportunities: Regular Research Grants
• FAPESP's process: 12/22274-2 - Study in the photochemical Wolff rearrangement from a,b-unsaturated diazoketones. Aplication in diversity synthesis of nitrogen and oxygen heterocycles
• Grantee: Barbara Bernardim de Souza
• Support Opportunities: Scholarships in Brazil - Doctorate