Evaluating the Catalytic Efficiency of the Human Membrane-type 1 Matrix Metalloproteinase (MMP-14) Using AuNP–Peptide Conjugates

• Published in: Journal of the American Chemical Society Vol. 145; no. 8; pp. 4570 - 4582
• Main Authors: Jin, Zhicheng, Dridi, Narjes, Palui, Goutam, Palomo, Valle, Jokerst, Jesse V., Dawson, Philip E., Amy Sang, Qing-Xiang, Mattoussi, Hedi
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.03.2023
• Subjects: Gold; Humans; Hydrolysis; Matrix Metalloproteinase 14; Metal Nanoparticles; Peptides
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/jacs.2c12032

Interactions of plasmonic nanocolloids such as gold nanoparticles and nanorods with proximal dye emitters result in efficient quenching of the dye photoluminescence (PL). This has become a popular strategy for developing analytical biosensors relying on this quenching process for signal transduction. Here, we report on the use of stable PEGylated gold nanoparticles, covalently coupled to dye-labeled peptides, as sensitive optically addressable sensors for determining the catalytic efficiency of the human matrix metalloproteinase-14 (MMP-14), a cancer biomarker. We exploit real-time dye PL recovery triggered by MMP-14 hydrolysis of the AuNP−peptide-dye to extract quantitative analysis of the proteolysis kinetics. Sub-nanomolar limit of detections for MMP-14 has been achieved using our hybrid bioconjugates. In addition, we have used theoretical considerations within a diffusion-collision framework to derive enzyme substrate hydrolysis and inhibition kinetics equations, which allowed us to describe the complexity and irregularity of enzymatic proteolysis of nanosurface-immobilized peptide substrates. Our findings offer a great strategy for the development of highly sensitive and stable biosensors for cancer detection and imaging.