Abstract
Protease-activated triggers have been introduced into a pore-forming protein, staphylococcal a-hemolysin (aHL). The hemolysin was remodeled by genetic engineering to form two-chain constructs with redundant polypeptide sequences at the central loop, the Integrity of which is crucial for efficient pore formation. The new hemolysins are activated when the polypeptide extensions are removed by proteases. By alterating the protease recognition sequence In the loop, selective activation by specified proteases can be obtained. Protease-triggered pore-forming proteins might be used for the selective destruction of cancer cells that bear tumor-associated proteases. When certain two-chain constructs are treated with proteases, a full-length polypeptide chain forms as the result of a protease-mediated transpeptidation reaction. This reaction might be used to produce chimeric hemolysins that are inaccessible by conventional routes.
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Acknowledgments
Supported by grants from the Department of Energy and the American Cancer Society (Massachusetts Division). Nathan Walsh is at Brown University and was supported at the Worcester Foundation by the National Science Foundation through the Research Experiences for Undergraduates Program.
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Walker, B., Walsh, N. & Bayley, H. Genetically-Engineered Protease-Activated Triggers in a Pore-Forming Protein. MRS Online Proceedings Library 330, 209–215 (1993). https://doi.org/10.1557/PROC-330-209
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DOI: https://doi.org/10.1557/PROC-330-209