Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

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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.

• Tipo de documento: Artigo Científico
• Fonte: NATURE COMMUNICATIONS; v. 7, OCT 26 2016.
• Citações Web of Science: 31

Resumo

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)

• Processo FAPESP: 15/07509-1 - Modificação específica de proteínas via reação de Horner-Wadsworth-Emmons seguida de rearranjo de Wolff
• Beneficiário: Barbara Bernardim de Souza
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 13/25504-1 - Desenvolvimento de novas metodologias para a-funcionalização assimétrica de compostos carbonílicos utilizando catalisadores quirais
• Beneficiário: Antonio Carlos Bender Burtoloso
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 12/22274-2 - Estudo do Rearranjo de Wolff fotoquímico a partir de diazocetonas a,b-insaturadas. Aplicação na síntese divergente de heterociclos de nitrogênio e oxigênio
• Beneficiário: Barbara Bernardim de Souza
• Modalidade de apoio: Bolsas no Brasil - Doutorado