Organotellurium Probes Enable One-step Single-cell Analysis of Post-translational Modification

Area of Science:

• Biochemistry
• Cell Biology
• Proteomics

Background:

• Protein prenylation is a crucial post-translational modification regulating cell signaling and membrane association.
• Dysregulation of protein prenylation is linked to various diseases.
• Current methods for studying prenylation involve multi-step metabolic labeling and bioorthogonal reactions.

Purpose of the Study:

• To develop a novel, single-step method for quantifying protein prenylation at the single-cell level.
• To apply this method to investigate prenylation changes in cellular models relevant to aging.
• To identify protein prenylation targets using chemical proteomics.

Main Methods:

• Development of tellurium-containing isoprenoid analogues for metabolic labeling.
• Application of mass cytometry for single-cell quantification of prenylation.
• Utilizing oxidation-controlled strain-promoted tellurophene-alkyne cycloaddition for target identification.

Main Results:

• A single-step enzymatic incorporation of tellurium-labeled analogues was achieved.
• Direct quantification of prenylation in single cells using mass cytometry was demonstrated.
• Perturbed prenylation levels were observed in autophagy-deficient L6 cells, a model for aging.
• Identification of prenylation targets through chemical proteomics was successful.

Conclusions:

• The developed tellurium-based methodology offers a robust and efficient approach for studying protein prenylation.
• This method enables direct, single-cell level analysis and target identification, bridging proteomic and multiplexed analyses.
• The findings open new avenues for investigating post-translational modifications and their roles in disease and aging.