Biotin methyl ester enhances cargo release in RUSH system and enables rapid biotinylation with TurboID

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Biology

Background:

• Biotin-streptavidin technology is crucial in biotechnology.
• Engineered streptavidin and biotin derivatives offer diverse research applications.

Purpose of the Study:

• To investigate biotin methyl ester (BME) as a novel biotin derivative.
• To evaluate BME's efficacy in cell penetration, the RUSH system, and TurboID biotinylation.

Main Methods:

• Utilized biotin methyl ester (BME) with a masked charged carboxyl group.
• Assessed BME's performance in the retention using selective hooks (RUSH) system.
• Measured biotinylation efficiency using the promiscuous biotin ligase TurboID.

Main Results:

• BME demonstrated improved cell penetration due to its masked carboxyl group.
• BME enabled prompt and uniform cargo release in the RUSH system.
• BME facilitated faster intracellular biotinylation compared to biotin with TurboID.
• BME acts as a biotin prodrug, enhancing RUSH and TurboID functionalities.

Conclusions:

• Biotin methyl ester (BME) offers significant advantages for biotin-streptavidin applications in living cells.
• BME enhances the RUSH system and TurboID-mediated biotinylation.
• BME holds potential as a biotin supplement for specific inherited metabolic disorders.