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Detection of Matrix Metalloproteinase Activity by Bioluminescence via Intein-Mediated Biotinylation of Luciferase

1,2,†, 1,†, 1,†, 1,2,†, 1,3,* and 1,2,3,4,*
1
Department of Life Science, Hanyang University, Seoul 04763, Korea
2
Brain Korea 21 Plus BioDefense Research Team, Hanyang University, Seoul 04763, Korea
3
Research Institute for Natural Sciences, Hanyang University, Seoul 04763, Korea
4
Research Institute for Convergence of Basic Sciences, Hanyang University, Seoul 04763, Korea
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Sensors 2018, 18(3), 875; https://doi.org/10.3390/s18030875
Received: 12 February 2018 / Revised: 13 March 2018 / Accepted: 14 March 2018 / Published: 15 March 2018
(This article belongs to the Special Issue Protein-Based Biosensors)
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Abstract

We report bioluminescence analysis of matrix metalloproteinase (MMP) activity in biological substances using a surface-bound luciferase probe. Intein-fused luciferase protein enables site-specific biotinylation of luciferase in the presence of N-terminus cysteine-biotin via intein-mediated splicing process, resulting in a strong association with high bioluminescence signal onto a NeutrAvidin-coated surface. When the peptide substrate for MMP-7 was inserted into a region between luciferase and intein, the biotinylated probe detected MMP-7 activity by cleaving the peptide, and surface-induced bioluminescence signal was strongly reduced in the MMP-secreted media or mouse tissue extracts, compared with that in MMP-deficient control set. Our approach is anticipated to be useful for generating biotinylated proteins and for their applications in diagnosing MMP activity in human diseases. View Full-Text
Keywords: luciferase; bioluminescence; matrix metalloproteinase; intein; biotinylation luciferase; bioluminescence; matrix metalloproteinase; intein; biotinylation
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Nguyen, D.L.; Kim, H.; Kim, D.; Lee, J.O.; Gye, M.C.; Kim, Y.-P. Detection of Matrix Metalloproteinase Activity by Bioluminescence via Intein-Mediated Biotinylation of Luciferase. Sensors 2018, 18, 875.

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