Xenoprotein engineering via synthetic libraries
Name
E5298.full.pdf
Size
1.62 MB
Format
Adobe PDF
Checksum (MD5)
75543ee2910615514d79a9fe1919fa62
Author(s)
Gates, Zachary P
Vinogradov, Alexander Alexandrovich
Quartararo, Anthony James
Bandyopadhyay, Anupam
Choo, Zi-Ning
Evans, Ethan Daniel
Halloran, Kathryn
Mijalis, Alexander James
Mong, Surin Khai
Simon, Mark
Date Issued
May 2018
Journal
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences (U.S.)
Citation
Gates, Zachary P. et al. “Xenoprotein Engineering via Synthetic Libraries.” Proceedings of the National Academy of Sciences 115, 23 (May 2018): E5298–E5306 © 2018 National Academy of Sciences
Version
Final published version
Abstract
Chemical methods have enabled the total synthesis of protein molecules of ever-increasing size and complexity. However, methods to engineer synthetic proteins comprising noncanonical amino acids have not kept pace, even though this capability would be a distinct advantage of the total synthesis approach to protein science. In this work, we report a platform for protein engineering based on the screening of synthetic one-bead one-compound protein libraries. Screening throughput approaching that of cell surface display was achieved by a combination of magnetic bead enrichment, flow cytometry analysis of on-bead screens, and high-throughput MS/MS-based sequencing of identified active compounds. Direct screening of a synthetic protein library by these methods resulted in the de novo discovery of mirror-image miniprotein-based binders to a ∼150-kDa protein target, a task that would be difficult or impossible by other means. Keywords: xenoprotein; mirror-image miniprotein; D-protein; protein engineering; flow cytometry
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
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.
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1073/PNAS.1722633115
