A short guide on blue fluorescent proteins: limits and perspectives
Pil-Won Seo1, Geun-Joong Kim2, Jeong-Sun Kim3
1Department of Chemistry, Chonnam National University, Gwangju, 61186, Republic of Korea.
Applied Microbiology and Biotechnology
|February 14, 2024
Summary
This review explores fluorescent proteins (FPs), comparing oxidation-dependent green fluorescent proteins (GFPs) with redox-free, NADPH-dependent blue fluorescent proteins (BFPs). NADPH-dependent BFPs show promise for future cellular imaging applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Fluorescent proteins (FPs) enable detection of macromolecules and metabolite abundance in organelles.
- Green fluorescent proteins (GFPs) and red fluorescent proteins (RFPs) rely on oxidative environments for chromophore formation.
- Cofactor-dependent proteins offer fluorescence in both aerobic and anaerobic conditions.
Purpose of the Study:
- To review GFP-derived and NADPH-dependent blue fluorescent proteins (BFPs).
- To highlight the potential of NADPH-dependent BFPs for future cellular applications.
- To discuss limitations of current FP technologies.
Main Methods:
- Comparative analysis of GFP-like and NADPH-dependent BFPs.
- Exploration of cofactor-dependent fluorescence mechanisms.
- Review of potential applications with advanced microscopy techniques.
Main Results:
- GFP-like FPs are limited to oxidative environments.
- NADPH-dependent BFPs function in aerobic and anaerobic conditions.
- Cofactor-dependent BFPs exhibit poor photostability and high background fluorescence.
Conclusions:
- NADPH-dependent BFPs offer a redox-free alternative to traditional FPs.
- Future advancements may involve coupling BFPs with two-photon fluorescence microscopy.
- Further development is needed to overcome photostability and background limitations in BFPs.
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