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Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

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Sensitive Method To Analyze Cell Surface GPI-Anchored Proteins Using DNA Hybridization Chain Reaction-Mediated Signal

Sayan Kundu¹, Kendall C Craig¹, Palak Gupta¹

¹Department of Chemistry, University of Florida, Gainesville, Florida 32611, United States.

Analytical Chemistry

|May 29, 2024

Summary

Researchers developed a novel method for detecting glycosylphosphatidylinositol-anchored proteins (GPI-APs) using metabolic engineering and DNA signal amplification. This technique enhances sensitivity for studying these vital cell surface proteins.

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Area of Science:

Biochemistry
Molecular Biology
Cell Biology

Background:

Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are crucial cell surface proteins.
Their low abundance poses significant challenges for detection and analysis.

Purpose of the Study:

To develop a novel, highly sensitive method for detecting and studying GPI-APs.
To overcome the limitations of existing techniques for analyzing low-abundance cell surface proteins.

Main Methods:

Metabolic engineering of cell surface GPI-APs using azido-inositol derivatives to introduce azido groups.
Coupling of modified GPI-APs with alkyne-functionalized DNA assemblies via hybridization chain reaction (HCR).
Utilizing DNA-facilitated fluorescence signal amplification for enhanced detection.

Main Results:

Significantly improved detection limits and sensitivity for GPI-APs.
Successful application in analyzing GPI-AP engineering and expression profiling.
Enabled investigation of live cells expressing GPI-APs.

Conclusions:

The developed method offers a powerful tool for advancing GPI-AP research.
This technique enhances the ability to study the function and dynamics of GPI-APs.
Facilitates broader biological investigations involving GPI-anchored proteins.