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Related Experiment Videos

A powerful combinatorial screen to identify high-affinity terbium(III)-binding peptides.

Mark Nitz1, Katherine J Franz, Rebecca L Maglathlin

  • 1Department of Chemistry Massachusetts Institute of Technology Cambridge, MA 02139, USA.

Chembiochem : a European Journal of Chemical Biology
|April 3, 2003
PubMed
Summary

Researchers developed a novel screening method to discover new lanthanide-binding tags (LBTs) with high affinity for Tb(3+) ions. This led to a 17-amino acid peptide with significantly enhanced binding and luminescence properties.

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Materials Science

Background:

  • Lanthanide-binding tags (LBTs) are crucial for applications requiring high-affinity lanthanide ion chelation.
  • Existing LBTs have limitations in binding affinity and luminescence intensity for certain applications.
  • Development of novel LBTs with improved properties is essential for advancing bioimaging and sensing technologies.

Purpose of the Study:

  • To develop and validate a new screening methodology for identifying high-affinity lanthanide-binding tags (LBTs).
  • To discover novel LBT sequences with enhanced affinity and luminescence for Tb(3+) ions.
  • To characterize the binding properties of newly identified LBTs.

Main Methods:

  • Utilized solid-phase split-and-pool combinatorial peptide synthesis for library generation.

Related Experiment Videos

  • Employed a two-step screening procedure involving photochemical release into an agarose matrix and aqueous solution evaluation.
  • Incorporated orthogonally cleavable linkers for efficient peptide separation and analysis.

    • Main Results:

    • Identified a linear 17-amino acid peptide sequence with a 140-fold increase in affinity for Tb(3+) (57 nM K(D)) compared to previous LBTs.
    • Macrocyclization of the linear peptide via a disulfide bond yielded a variant with a 2-nM apparent dissociation constant for Tb(3+).
    • The new LBTs exhibit intense luminescence properties suitable for various applications.

    Conclusions:

    • The developed screening methodology is effective for discovering high-affinity lanthanide-binding peptides.
    • The novel linear and macrocyclic LBTs represent significant advancements in lanthanide ion binding and luminescence.
    • These findings open new avenues for developing advanced biosensors, imaging agents, and other lanthanide-based technologies.