Dual-Product Synergistically Enhanced Colorimetric Assay for Sensitive Detection of Lipid Transferase Activity

Area of Science:

• Biochemistry
• Nanotechnology
• Analytical Chemistry

Background:

• Protein lipidation, catalyzed by lipid transferases, is crucial for physiological processes.
• Lipid transferases are increasingly recognized as therapeutic targets for diseases such as cancer and neurodegeneration.
• Sensitive detection of lipid transferase activity in biological samples remains a significant challenge.

Purpose of the Study:

• To develop a sensitive and convenient colorimetric method for detecting lipid transferase activity.
• To utilize gold nanoparticles (AuNPs) with a dual-product synergistic enhancement strategy for improved sensitivity.
• To establish a visual detection platform for lipid transferase activity, applicable to drug screening and biological sample analysis.

Main Methods:

• A gold nanoparticle (AuNP)-based colorimetric assay was designed.
• The assay utilizes substrate peptides for Homo sapiens N-myristoyltransferase 1 (HsNMT1) and monitors changes in AuNP aggregation.
• Detection relies on charge reversal of biomolecules and formation of Au-S bonds, leading to color changes in AuNPs due to altered surface plasmon resonance.

Main Results:

• The developed method achieved sensitive detection of HsNMT1 activity with a linear range of 2-75 nM and a low detection limit of 0.56 nM.
• The assay provided a visual readout of enzyme activity through naked-eye observation of color changes in AuNPs.
• The method was successfully applied to analyze HsNMT1 activity in cell lysates and for inhibitor screening.

Conclusions:

• The proposed AuNP-based colorimetric assay offers a sensitive and convenient platform for detecting lipid transferase activity.
• The assay's design, leveraging dual-product synergistic enhancement, shows promise for universal application to various lipid transferases.
• This method holds significant potential for lipid transferase-targeted drug development and screening.