Preparation of unnatural N-to-N and C-to-C protein fusions
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Witte-2012-Preparation of unnat.pdf
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Author(s)
Witte, Martin D.
Cragnolini, Juan J.
Dougan, Stephanie K.
Yoder, Nicholas C.
Ploegh, Hidde
Popp, Maximilian W.L.
Date Issued
July 2012
Journal
Proceedings of the National Academy of Sciences of the United States of America
Publisher
National Academy of Sciences (U.S.)
Citation
Witte, M. D. et al. “Preparation of Unnatural N-to-N and C-to-C Protein Fusions.” Proceedings of the National Academy of Sciences 109.30 (2012): 11993–11998. Web. © 2012 National Academy of Sciences.
Version
Final published version
Abstract
Standard genetic approaches allow the production of protein composites by fusion of polypeptides in head-to-tail fashion. Some applications would benefit from constructions that are genetically impossible, such as the site-specific linkage of proteins via their N or C termini, when a remaining free terminus is required for biological activity. We developed a method for the production of N-to-N and C-to-C dimers, with full retention of the biological activity of both fusion partners and without inflicting chemical damage on the proteins to be joined. We use sortase A to install on the N or C terminus of proteins of interest the requisite modifications to execute a strain-promoted copper-free cycloaddition and show that the ensuing ligation proceeds efficiently. Applied here to protein–protein fusions, the method reported can be extended to connecting proteins with any entity of interest.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Whitehead Institute for Biomedical Research
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1073/pnas.1205427109
