Oligo-Asp Tag/Zn(II) Complex Probe as a New Pair for Labeling and Fluorescence Imaging of Proteins

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,...

Full description

Saved in:
Bibliographic Details
Published in:Journal of the American Chemical Society 2006-08, Vol.128 (32), p.10452-10459
Main Authors: Ojida, Akio, Honda, Kei, Shinmi, Daisuke, Kiyonaka, Shigeki, Mori, Yasuo, Hamachi, Itaru
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Aspartic Acid - chemistry
Aspartic Acid - metabolism
Calcium - analysis
Chemistry
CHO Cells
Circular Dichroism
Cricetinae
Cricetulus
Exact sciences and technology
Fluorescent Dyes - chemistry
General and physical chemistry
Models, Molecular
Molecular Sequence Data
Organic chemistry
Organometallic Compounds - chemical synthesis
Organometallic Compounds - chemistry
Peptides
Preparations and properties
Proteins - analysis
Thermodynamics
Zinc - chemistry
Zinc - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja0618604