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Related Experiment Videos

Engineering fluorescent proteins.

Atsushi Miyawaki1, Takeharu Nagai, Hideaki Mizuno

  • 1Laboratory for Cell Function Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2-1 Hirosawa,Wako-city, Saitama, 351-0198 Japan. matsushi@brain.riken.jp

Advances in Biochemical Engineering/Biotechnology
|August 6, 2005
PubMed
Summary

Researchers are advancing bright, fast-maturing fluorescent proteins, including novel photoactivatable and monomeric variants. These engineered proteins, like the green fluorescent protein (GFP), are crucial for developing sensitive biosensors for physiological signals.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biophysics

Background:

  • Green fluorescent protein (GFP) from Aequorea victoria and related Anthozoa proteins possess intrinsic chromophores.
  • Recent advancements focus on improving protein brightness and chromophore development efficiency.

Purpose of the Study:

  • To engineer novel fluorescent proteins with enhanced properties, including photoactivation, photoconversion, and monomeric forms.
  • To develop sensitive fluorescent probes for monitoring physiological signals like membrane potential and calcium concentrations.
  • To further characterize protein structure and maturation for rational design of improved variants.

Main Methods:

  • Engineering of bright, fast-maturing GFP variants.
  • Development of photoactivatable and photoconvertible fluorescent proteins.

Related Experiment Videos

  • Creation of monomeric fluorescent proteins for fusion applications.
  • Design of circularly permuted GFP variants for biosensing.

    • Main Results:

    • Obtained brighter fluorescent protein variants with rapid chromophore maturation.
    • Developed novel fluorescent proteins capable of photoactivation and photoconversion.
    • Engineered monomeric versions suitable for fusion protein applications.
    • Created fluorescent probes based on GFP and circularly permuted variants for sensing membrane potential and calcium.

    Conclusions:

    • Engineered fluorescent proteins offer improved fluorescence properties and functionality.
    • These advanced proteins are valuable tools for biosensing and molecular imaging.
    • Ongoing research into structure-maturation relationships will enable further rational design of tailored fluorescent proteins.