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

• Main Authors: Duc Long Nguyen, Hanim Kim, Dasom Kim, Jin Oh Lee, Myung Chan Gye, Young-Pil Kim
• Format: Article
• Language: English
• Published: MDPI AG 2018-03-01
• Series: Sensors
• Subjects: luciferase; bioluminescence; matrix metalloproteinase; intein; biotinylation
• Online Access: http://www.mdpi.com/1424-8220/18/3/875

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.