| Info. | Vol.4 - No.3 (2010.09.20) |
|---|---|
| Title | On-chip assay of matrix metalloproteinase-3 activity using fluorescence-conjugated gelatin arrays |
| Authors | Deok-Hoon Kong1 , Se-Hui Jung1 , Seung-Taek Lee2 & Kwon-Soo Ha1 |
| Institutions | 1Department of Molecular and Cellular Biochemistry,
Kangwon National University School of Medicine, Chuncheon,
Gangwon-do 200-701, Korea 2Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749, Korea Correspondence and requests for materials should be addressed t K.-S. Ha (ksha@kangwon.ac.kr) |
| Abstract | We developed a new high-throughput approach to measure matrix metalloproteinase (MMP)-3 activity using Cy3-conjugated gelatin arrays. We fabricated Cy3-conjugated gelatin arrays by immobilizing Cy3-conjugated gelatin onto well-type amine arrays. The ratio of Cy3-NHS to gelatin was optimized to improve the efficiency of the arrays. MMP-3 activity was determined by the decrease of fluorescence intensity due to MMP-3-mediated degradation of Cy3-conjugated gelatin. The gelatinolytic activity of MMP-3 increased in the presence of Brij-35, a non-ionic detergent, in a dose-dependent manner. Cy3-conjugated gelatin arrays were successfully applied for the analyses of MMP-3 activity, which showed a dose-dependent degradation of the Cy3-conjugated substrate. This chip-based activity assay using a Cy3-conjugated gelatin array is simple, rapid, cost-effective, and has a potential for activity-based proteomic researches and screening of MMP-3 inhibitors. |
| Keyword | Matrix metalloproteinase-3, Well-type amine arrays, On-chip activity assay, Cy3-conjugated gelatin arrays, High-throughput |
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