Incorporation of glycosylated amino acid into protein by an in vitro translation system

Teruhiko Matsubara, Kazutoshi Iijima, Takayoshi Watanabe, Takahiro Hohsaka, Toshinori Sato

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

O-GalNAcα-modified proteins are the precursor of mucin-type O-glycosylated proteins. Homogeneously O-glycosylated proteins are required to investigate the biological functions of glycoproteins and to develop biopharmaceuticals. Here we show that the incorporation of GalNAcα-Thr into proteins successfully proceeded by the use of a chemically aminoacylated tRNA. GalNAcα-Thr was chemoenzymatically attached to amber suppressor tRNA and the product was subjected to in vitro translation together with streptavidin mRNA containing the UAG codon. Gel electrophoresis and mass analysis showed that GalNAcα-Thr was successfully incorporated into the N-terminus, although it was not incorporated at the interior. This method will facilitate the preparation of homogeneous GalNAcα-proteins.

Original languageEnglish
Pages (from-to)5634-5636
Number of pages3
JournalBioorganic and Medicinal Chemistry Letters
Volume23
Issue number20
DOIs
Publication statusPublished - 2013 Oct 15

Keywords

  • Expanding genetic code
  • Glycoprotein
  • Glycosylated amino acid
  • In vitro translation system
  • O-Glycan

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

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