Sortase-catalyzed transformations that improve the properties of cytokines
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Popp-2011-Feb-Sortase-catalyzed tr.pdf
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Author(s)
Dougan, Stephanie K.
Chuang, Tzu-Ying
Spooner, Eric
Ploegh, Hidde
Popp, Maximilian W.L.
Date Issued
February 2011
Journal
Proceedings of the National Academy of Sciences of the United States of America
Publisher
National Academy of Sciences (U.S.)
Citation
Popp, M. W. et al. “Sortase-catalyzed transformations that improve the properties of cytokines.” Proceedings of the National Academy of Sciences 108 (2011): 3169-3174. ©2011 by the National Academy of Sciences.
Version
Final published version
Abstract
Recombinant protein therapeutics often suffer from short circulating half-life and poor stability, necessitating multiple injections and resulting in limited shelf-life. Conjugation to polyethylene glycol chains (PEG) extends the circulatory half-life of many proteins, but the methods for attachment often lack specificity, resulting in loss of biological activity. Using four-helix bundle cytokines as an example, we present a general platform that uses sortase-mediated transpeptidation to facilitate site-specific attachment of PEG to extend cytokine half-life with full retention of biological activity. Covalently joining the N and C termini of proteins to obtain circular polypeptides, again executed using sortase, increases thermal stability. We combined both PEGylation and circularization by exploiting two distinct sortase enzymes and the use of a molecular suture that allows both site-specific PEGylation and covalent closure. The method developed is general, uses a set of easily accessible reagents, and should be applicable to a wide variety of proteins, provided that their termini are not involved in receptor binding or function.
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.1016863108
