Measuring in vivo protein half-life

Chem Biol. 2011 Jun 24;18(6):805-15. doi: 10.1016/j.chembiol.2011.03.014.

Abstract

Protein 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD4 Antigens / chemistry
  • CD4 Antigens / genetics
  • CD4 Antigens / metabolism
  • Cells, Cultured
  • Female
  • Fibroblasts / metabolism
  • Fluorescent Dyes / chemistry
  • Half-Life
  • Mice
  • Mice, Nude
  • Mice, Transgenic
  • O(6)-Methylguanine-DNA Methyltransferase / chemistry
  • O(6)-Methylguanine-DNA Methyltransferase / genetics
  • O(6)-Methylguanine-DNA Methyltransferase / metabolism*
  • Protein Stability*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • CD4 Antigens
  • Fluorescent Dyes
  • Recombinant Fusion Proteins
  • O(6)-Methylguanine-DNA Methyltransferase