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Related Experiment Video

Updated: May 12, 2026

Directed Assembly of Elastin-like Proteins into defined Supramolecular Structures and Cargo Encapsulation In Vitro
10:01

Directed Assembly of Elastin-like Proteins into defined Supramolecular Structures and Cargo Encapsulation In Vitro

Published on: April 8, 2020

Engineering fluorogen activating proteins into self-assembling materials.

Matthew J Saunders1, Wen Liu, Christopher Szent-Gyorgyi

  • 1Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, PA, USA. matsaun@andrew.cmu.edu

Bioconjugate Chemistry
|April 12, 2013
PubMed
Summary
This summary is machine-generated.

A new bifunctional protein, dL5_EAK, combines a fluorogen activating protein (FAP) with an amphiphilic peptide for solid-phase fluorescence detection. This conjugate forms stable membranes, enhancing fluorescence for potential in situ biosensing applications.

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Published on: April 27, 2018

Area of Science:

  • Bioconjugation Chemistry
  • Materials Science
  • Biotechnology

Background:

  • Fluorogen activating proteins (FAPs) like dL5 enhance fluorescence of dyes such as malachite green (MG).
  • Amphiphilic peptides, such as AEAEAKAK, can self-assemble into structures like hydrogels.
  • Combining FAPs with self-assembling peptides could create novel detection platforms.

Purpose of the Study:

  • To develop a novel bifunctional protein, dL5_EAK, by conjugating the FAP dL5 with an amphiphilic peptide.
  • To investigate the coassembly of dL5_EAK with the self-assembling peptide EAK16-II into insoluble membrane composites.
  • To assess the fluorescence properties and stability of the resulting membrane composites for potential biosensing applications.

Main Methods:

  • Synthesis and characterization of the dL5_EAK conjugate.
  • Coassembly studies with EAK16-II using denaturing polyacrylamide electrophoresis.
  • Fluorescence intensity measurements of the membrane composites.
  • Scanning electron microscopy (SEM) for structural analysis.

Main Results:

  • Greater than 78% of dL5_EAK successfully incorporated into EAK16-II membranes, unlike dL5 alone.
  • Membranes containing dL5_EAK showed a 4-fold increase in fluorescence intensity compared to dL5/EAK16-II mixtures.
  • SEM revealed the presence of FAP particulates integrated within the membrane fibrils.
  • The FAP's fluorescence-enhancing capability remained intact within the composite structure.

Conclusions:

  • The dL5_EAK conjugate effectively coassembles with EAK16-II to form stable, fluorescent membrane composites.
  • This system demonstrates potential for developing in situ forming local sensors for detecting analytes like MG.
  • The immobilized FAP in coacervate membranes could enable monitoring of biological processes, such as inflammatory cell migration.