Semisynthesis of a protein with cholesterol at the c-terminal, targeted to the cell membrane of live cells

Kenta Teruya, Keiko Nishizawa, Katsumi Doh-Ura

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

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 languageEnglish
Pages (from-to)493-500
Number of pages8
JournalProtein Journal
Volume29
Issue number7
DOIs
Publication statusPublished - 2010 Oct

Keywords

  • C-terminal
  • Cell membrane
  • Chemical modification
  • Cholesterol
  • Semi-synthesis

ASJC Scopus subject areas

  • Analytical Chemistry
  • Bioengineering
  • Biochemistry
  • Organic Chemistry

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