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On-chip assay of matrix metalloproteinase-3 activity using fluorescence-conjugated gelatin arrays

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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.

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Authors and Affiliations

  1. Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine, Chuncheon, Gangwon-do, 200-701, Korea

    Deok-Hoon Kong, Se-Hui Jung & Kwon-Soo Ha

  2. Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul, 120-749, Korea

    Seung-Taek Lee

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  1. Deok-Hoon Kong
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  2. Se-Hui Jung
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  4. Kwon-Soo Ha
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Correspondence to Kwon-Soo Ha.

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Kong, DH., Jung, SH., Lee, ST. et al. On-chip assay of matrix metalloproteinase-3 activity using fluorescence-conjugated gelatin arrays. BioChip J 4, 210–216 (2010). https://doi.org/10.1007/s13206-010-4308-4

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  • DOI: https://doi.org/10.1007/s13206-010-4308-4

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