Quantitation of Protein Translation Rate In Vivo with Bioorthogonal Click-Chemistry.
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.
Instituto de Biología Molecular y Celular de Plantas, CSIC-Universidad Politécnica de Valencia, Valencia, Spain.
Borja
B
Belda-Palazón
Instituto de Biología Molecular y Celular de Plantas, CSIC-Universidad Politécnica de Valencia, Valencia, Spain.
Alejandro
A
Ferrando
Centro de Investigación Príncipe Felipe, Valencia, Spain. rfarras@cipf.es.
Rosa
R
Farràs
D015153:Blotting, Western; D057930:Click Chemistry / Q000379:methods*; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
D011506:Proteins
Affinity purification; Bioorthogonal amino acid; Click-chemistry; Translation rate; Western blot