Diversification of polyphosphate end-labeling via bridging molecules

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Inorganic polyphosphate's biological roles are under investigation.
• Previous methods allowed covalent coupling of amine-containing molecules to polyphosphate terminals.
• Further development is needed to expand labeling capabilities.

Purpose of the Study:

• To optimize and extend a carbodiimide-based method for labeling inorganic polyphosphate.
• To enable the attachment of diverse molecules to polyphosphate terminals.
• To develop efficient purification strategies for labeled polyphosphate.

Main Methods:

• Optimized carbodiimide reaction conditions for polyphosphate labeling.
• Utilized bridging molecules with primary amine and additional reactive groups (thiol, click chemistry).
• Developed three facile purification methods for conjugated polyphosphate.

Main Results:

• Successfully labeled inorganic polyphosphate with biotin, peptide epitope tags, and fluorescent dyes.
• Demonstrated a versatile two-step labeling approach.
• Established efficient purification techniques for labeled polyphosphate.

Conclusions:

• The optimized method provides a versatile platform for labeling inorganic polyphosphate.
• This facilitates further investigation into the biological functions of polyphosphate.
• The developed purification methods ensure high-quality labeled products.