Oligo-Asp Tag/Zn(II) complex probe as a new pair for labeling and fluorescence imaging of proteins

Akio Ojida, Kei Honda, Daisuke Shinmi, Shigeki Kiyonaka, Yasuo Mori, Itaru Hamachi

Research output: Contribution to journalArticle

139 Citations (Scopus)

Abstract

To accomplish the selective labeling of a specific protein in complicated biological systems, a peptide tag incorporated into the protein and a complementary small molecular probe are required. Although a variety of peptide tag/probe pairs have been developed as molecular tools for protein analyses, the availability of pairs suitable for real-time imaging of proteins is still limited. We now report a new peptide tag/artificial probe pair composed of a genetically encodable oligo-aspartate sequence (D4 tag, (D4)n, n = 1-3) and the corresponding multinuclear Zn(II) complexes (Zn(II)-DpaTyrs). The strong binding affinity of the Zn(II)-DpaTyr probes with the D4 tag was a result of the multiple coordination bonds and the multivalent effect. It was measured quantitatively by isothermal titration calorimetry. The high affinity between the tag and the probe, indispensable for the selective protein labeling, enabled the pair to be used for the labeling and fluorescence imaging of a membrane-bound receptor protein tethering a triply repeated D4 tag ((D4) 3) in an intact cell configuration without significantly affecting the receptor signal transduction.

Original languageEnglish
Pages (from-to)10452-10459
Number of pages8
JournalJournal of the American Chemical Society
Volume128
Issue number32
DOIs
Publication statusPublished - Aug 16 2006
Externally publishedYes

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Optical Imaging
Labeling
Fluorescence
Proteins
Imaging techniques
Peptides
Molecular Probes
Signal transduction
Calorimetry
Biological systems
Titration
Aspartic Acid
Signal Transduction
Availability
Membranes

All Science Journal Classification (ASJC) codes

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

Cite this

Oligo-Asp Tag/Zn(II) complex probe as a new pair for labeling and fluorescence imaging of proteins. / Ojida, Akio; Honda, Kei; Shinmi, Daisuke; Kiyonaka, Shigeki; Mori, Yasuo; Hamachi, Itaru.

In: Journal of the American Chemical Society, Vol. 128, No. 32, 16.08.2006, p. 10452-10459.

Research output: Contribution to journalArticle

Ojida, Akio ; Honda, Kei ; Shinmi, Daisuke ; Kiyonaka, Shigeki ; Mori, Yasuo ; Hamachi, Itaru. / Oligo-Asp Tag/Zn(II) complex probe as a new pair for labeling and fluorescence imaging of proteins. In: Journal of the American Chemical Society. 2006 ; Vol. 128, No. 32. pp. 10452-10459.
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