Detection of Matrix Metalloproteinase Activity by Bioluminescence via Intein-Mediated Biotinylation of Luciferaseopen access
- Authors
- Nguyen, Duc Long; Kim, Hanim; Kim, Dasom; Lee, Jin Oh; Gye, Myung Chan; Kim, Young-Pil
- Issue Date
- Mar-2018
- Publisher
- MDPI
- Keywords
- luciferase; bioluminescence; matrix metalloproteinase; intein; biotinylation
- Citation
- SENSORS, v.18, no.3
- Indexed
- SCIE
SCOPUS
- Journal Title
- SENSORS
- Volume
- 18
- Number
- 3
- URI
- https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/17736
- DOI
- 10.3390/s18030875
- ISSN
- 1424-8220
- 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.
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