The high performance of 3XFLAG for target purification of a bioactive metabolite: A tag combined with a highly effective linker structure

Minoru Ueda, Yoshiyuki Manabe, Makoto Mukai

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Affinity purification using immunoprecipitation (IP) is an extremely useful method for target profiling of bioactive natural products. We examined IP purification of CMetE, which is a molecular target for potassium isolespedezate (1), a leaf-opening factor of Cassia plant. We studied IP efficiency using a panel of FLAG-connected molecular probes (2-8), including probes with varying structures and lengths of the linker moiety. The results suggest that not only the length, but the chemical nature of the linker moiety, strongly affect the IP efficiency. 3XFLAG, a tag combined with a linker moiety of charged amino acids, gave the best results and was most useful for IP purification of the molecular target.

Original languageEnglish
Pages (from-to)1359-1362
Number of pages4
JournalBioorganic and Medicinal Chemistry Letters
Volume21
Issue number5
DOIs
Publication statusPublished - 2011 Mar 1

Keywords

  • Immunoprecipitation
  • Linker
  • Molecular probe
  • Target protein

ASJC Scopus subject areas

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

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