An engineered monomeric Zoanthus sp. yellow fluorescent protein

Area of Science:

• Biochemistry
• Molecular Biology
• Biophysics

Background:

• Monomeric fluorescent proteins (FPs) are crucial tools in molecular biology.
• A spectral gap exists between red-shifted green FP (GFP) and blue-shifted red FP (RFP) variants.
• There is a need for novel FPs to expand multicolor imaging and biosensing capabilities.

Purpose of the Study:

• To engineer a monomeric yellow fluorescent protein (YFP) to fill the spectral gap between existing FP variants.
• To characterize the properties of the engineered YFP, named mPapaya1.
• To evaluate mPapaya1's utility in Förster resonance energy transfer (FRET) applications.

Main Methods:

• Protein engineering of a yellow FP from Zoanthus sp. coral.
• Characterization of mPapaya1's brightness, photostability, and monomeric nature in vitro and in living cells.
• Demonstration of mPapaya1 as a FRET acceptor with mTFP1 donor.

Main Results:

• Successfully engineered a monomeric YFP variant, mPapaya1.
• mPapaya1 exhibits excellent fluorescent brightness and good photostability.
• mPapaya1 maintains its monomeric character in vitro and in cellular contexts.
• mPapaya1 functions effectively as a FRET acceptor when paired with mTFP1.

Conclusions:

• mPapaya1 effectively bridges the spectral gap in the FP palette.
• mPapaya1 is a valuable tool for multicolor imaging and FRET-based biosensing.
• The engineered mPapaya1 expands the available repertoire of fluorescent proteins for biological research.