Measuring In Vivo Protein Half-Life
| Autor: | Didier Trono, Sandra Offner, Karolina Bojkowska, Isabelle Barde, Sonia Verp, Christian Heinis, Francesca R. Santoni de Sio, Kai Johnsson |
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| Rok vydání: | 2011 |
| Předmět: |
Noninvasive imaging
Tag Fusion Proteins Extracellular proteins Living Cells Recombinant Fusion Proteins Clinical Biochemistry Mice Nude Expression Mice Transgenic Biology Superresolution 010402 general chemistry 01 natural sciences Biochemistry Cd4 Degradation Mice O(6)-Methylguanine-DNA Methyltransferase 03 medical and health sciences Ubiquitin In vivo Lentiviral Vectors O-6-Alkylguanine-Dna Alkyltransferase Drug Discovery Animals Molecular Biology Cells Cultured Fluorescent Dyes 030304 developmental biology Pharmacology 0303 health sciences Protein Stability Protein turnover General Medicine Fibroblasts Fusion protein 0104 chemical sciences Cell biology O-6-Benzylguanine CD4 Antigens biology.protein Molecular Medicine Female Intracellular Chemical labeling Half-Life |
| Zdroj: | Chemistry & biology |
| ISSN: | 1074-5521 |
| DOI: | 10.1016/j.chembiol.2011.03.014 |
| Popis: | SummaryProtein turnover critically influences many biological functions, yet methods have been lacking to assess this parameter in vivo. Here, we demonstrate how chemical labeling of SNAP-tag fusion proteins can be exploited to measure the half-life of resident intracellular and extracellular proteins in living mice. First, we demonstrate that SNAP-tag substrates have wide bioavailability in mice and can be used for the specific in vivo labeling of SNAP-tag fusion proteins. We then apply near-infrared probes to perform noninvasive imaging of in vivo-labeled tumors. Finally, we use SNAP-mediated chemical pulse-chase labeling to perform measurement of the in vivo half-life of different extra- and intracellular proteins. These results open broad perspectives for studying protein function in living animals. |
| Databáze: | OpenAIRE |
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