All relevant data are within the paper and its Supporting Information files.įunding: This work was supported by a Grant-in-Aid for Scientific Research on Innovative Areas "Integrative Research on Cancer Microenvironment Network" from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to SKK) and partially supported by Grants-in-Aid from the Japan Society for Promotion of Science 22700469 (to T. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: The authors confirm that all data underlying the findings are fully available without restriction. Received: ApAccepted: JPublished: August 1, 2014Ĭopyright: © 2014 Kadonosono et al. PLoS ONE 9(8):Įditor: Jody Michael Mason, University of Essex, United Kingdom (2014) A Fluorescent Protein Scaffold for Presenting Structurally Constrained Peptides Provides an Effective Screening System to Identify High Affinity Target-Binding Peptides. These results demonstrate that gFPS has the potential to serve as a powerful tool to improve screening of structurally constrained peptides that have a high target affinity, and suggest that it could expedite the one-step identification of clinically applicable cancer cell-binding peptides.Ĭitation: Kadonosono T, Yabe E, Furuta T, Yamano A, Tsubaki T, Sekine T, et al. In addition, gFPS could be expressed on the yeast cell surface and applied for a high-throughput screening. When gFPSs presenting human epidermal growth factor receptor type 2 (HER2)-targeting peptides were added to the culture medium of HER2-expressing cells, we could easily identify the peptides with high HER2-affinity and -specificity based on gFPS fluorescence. gFPS can present 4 to 12 exogenous amino acids without a loss of fluorescence. Molecular dynamics simulation supported the suitability of this site for presentation of exogenous peptides with a constrained structure. To efficiently identify high-affinity target-binding peptides, we have constructed a fluorescent protein scaffold, designated gFPS, in which structurally constrained peptides are integrated at residues K131–L137 of superfolder green fluorescent protein. However, unstructured peptides often fail to bind their target molecules with high affinity. Peptides that have high affinity for target molecules on the surface of cancer cells are crucial for the development of targeted cancer therapies.
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