Abstract
Human mast cells contain proteases that are important functional components and serve as markers of mast cell activation or degranulation. Although tryptase is the best recognized mast cell protease, chymase and Cathepsin G also are found in some human mast cells. Methods for measuring the activities of these enzymes using sensitive synthetic peptide thiobenzyl ester substrates are described in this chapter. Using a visible plate reader with kinetic software femtomole quantities of these proteases can be measured. These methods are demonstrated in assays of tryptase and chymase in cell-free extracts of the HMC-1 and 5C6 human mast cell lines, as well as in extracts of cord blood derived mast cells.
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Acknowledgments
The author is grateful for the assistance of Andrew Hull and Susannah Johnson, who performed portions of this work as part of their undergraduate research experiences. Appreciation is expressed to Dr. James Powers (Georgia Tech.) for introducing the author to thiolbenzyl ester substrates. This work was supported by NIH grant R15 AI45549.
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Johnson, D.A. (2006). Human Mast Cell Proteases. In: Krishnaswamy, G., Chi, D.S. (eds) Mast Cells. Methods in Molecular Biology, vol 315. Humana Press. https://doi.org/10.1385/1-59259-967-2:193
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DOI: https://doi.org/10.1385/1-59259-967-2:193
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