Quantitation of Protein Translation Rate In Vivo with Bioorthogonal Click-Chemistry

Autores de CIPF

• 

  Rosa M Farràs Rivera

  Autor

Participantes ajenos a CIPF

• Belda-Palazon, B
• Ferrando, A

Grupos de Investigación

• Laboratorio de Señalización Oncogénica

  

  Jefe/a de Grupo

  Rosa M Farràs Rivera

Abstract

The development of novel bioorthogonal reactives that can be used to tag biomolecules in vivo has revolutionized the studies of cellular and molecular biology. Among those novel reactive substances, amino acid analogs can be used to label nascent proteins, thus opening new avenues for measuring protein translation rates in vivo with a limited manipulation of the sample. Here, we describe the use of Click-chemistry to tag and separate newly synthesized proteins in mammalian cells that can be used, coupled with western analysis, to estimate the translation rate of any protein of interest.

Keywords

• Click-chemistry; Bioorthogonal amino acid; Translation rate; Affinity purification; Western blot