Abstract
Various proteins are modified post-translationally to localize them at the cell membrane. Among them, hedgehog-family proteins are modified by cholesterol at the C-terminal. In this study, green fluorescent protein (GFP) modified with cholesterol (GFP-Chol) at the C-terminal was prepared semisynthetically and investigated. This semi-synthesis was performed using the following native chemical ligation: GFP-Cα-thioester was prepared using the intein-mediated thioester exchange reaction and was ligated to Cys-NH-diethylene glycol-NHCO-cholesterol in the presence of a detergent. After removal of the detergent, the GFP-Chol was applied to mouse live cells. Confocal laser fluorescent microscopy confirmed localization of GFP-Chol at the cell membrane. The findings suggest that modifying proteins with cholesterol at the C-terminal is useful for targeting the proteins to the cell membrane of live cells.
| Original language | English |
|---|---|
| Pages (from-to) | 493-500 |
| Number of pages | 8 |
| Journal | Protein Journal |
| Volume | 29 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 2010 Oct |
Keywords
- C-terminal
- Cell membrane
- Chemical modification
- Cholesterol
- Semi-synthesis
ASJC Scopus subject areas
- Analytical Chemistry
- Bioengineering
- Biochemistry
- Organic Chemistry