Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

• Tipo: Artigo em Revista Científica Internacional
• Data: 2016

Autores

Bernardim, B (Autor) Outra; Cal, PMSD (Autor) Outra; Matos, MJ (Autor) Outra; Oliveira, BL (Autor) Outra; Martinez Saez, N (Autor) Outra; Albuquerque, IS (Autor) Outra; Perkins, E (Autor) Outra; Corzana, F (Autor) Outra; Burtoloso, ACB (Autor) Outra; Jimenez Oses, G (Autor) Outra; Bernardes, GJL (Autor) Outra

Revista

Título: Nature Communications

Vol. 7

ISSN: 2041-1723

Editora: Springer Nature

Indexação

ISI Web of Knowledge - 62 Citações

Outras Informações

ID Authenticus: P-00M-626

DOI: 10.1038/ncomms13128

Abstract (EN): 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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 9