No-wash protein labeling with designed fluorogenic probes and application to real-time pulse-chase analysis

Shin Mizukami, Shuji Watanabe, Yuri Akimoto, Kazuya Kikuchi

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

Small molecule labeling techniques for cellular proteins under physiological conditions are very promising for revealing new biological functions. We developed a no-wash fluorogenic labeling system by exploiting fluorescence resonance energy transfer (FRET)-based fluorescein-cephalosporin- azopyridinium probes and a mutant β-lactamase tag. Fast quencher elimination, hydrophilicity, and high resistance against autodegradation were achieved by rational refinement of the structure. By applying the probe to real-time pulse-chase analysis, the trafficking of epidermal growth factor receptors between cell surface and intracellular region was imaged. In addition, membrane-permeable derivatization of the probe enabled no-wash fluorogenic labeling of intracellular proteins.

Original languageEnglish
Pages (from-to)1623-1629
Number of pages7
JournalJournal of the American Chemical Society
Volume134
Issue number3
DOIs
Publication statusPublished - 2012 Jan 25
Externally publishedYes

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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