Covalent Modification of Synthetic Hydrogels with Bioactive Proteins via Sortase-Mediated Ligation
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Griffith_Covalent modification.pdf
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Author(s)
Cambria, Elena
Kroll, Carsten
Krueger, Andrew T.
Imperiali, Barbara
Chopko Ahrens, Caroline
Cook, Christi Dionne
Renggli-Frey, Kasper
Griffith, Linda G
Date Issued
June 2015
Journal
Biomacromolecules
Publisher
American Chemical Society (ACS)
Citation
Cambria, Elena, Kasper Renggli, Caroline C. Ahrens, Christi D. Cook, Carsten Kroll, Andrew T. Krueger, Barbara Imperiali, and Linda G. Griffith. “Covalent Modification of Synthetic Hydrogels with Bioactive Proteins via Sortase-Mediated Ligation.” Biomacromolecules 16, no. 8 (August 10, 2015): 2316–26. © 2015 American Chemical Society
Version
Final published version
Abstract
Synthetic extracellular matrices are widely used in regenerative medicine and as tools in building in vitro physiological culture models. Synthetic hydrogels display advantageous physical properties, but are challenging to modify with large peptides or proteins. Here, a facile, mild enzymatic postgrafting approach is presented. Sortase-mediated ligation was used to conjugate human epidermal growth factor fused to a GGG ligation motif (GGG-EGF) to poly(ethylene glycol) (PEG) hydrogels containing the sortase LPRTG substrate. The reversibility of the sortase reaction was then exploited to cleave tethered EGF from the hydrogels for analysis. Analyses of the reaction supernatant and the postligation hydrogels showed that the amount of tethered EGF increases with increasing LPRTG in the hydrogel or GGG-EGF in the supernatant. Sortase-tethered EGF was biologically active, as demonstrated by stimulation of DNA synthesis in primary human hepatocytes and endometrial epithelial cells. The simplicity, specificity, and reversibility of sortase-mediated ligation and cleavage reactions make it an attractive approach for modification of hydrogels.
MIT Department
Massachusetts Institute of Technology. Center for Gynepathology Research
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemical Engineering
Massachusetts Institute of Technology. Department of Chemistry
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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/10.1021/acs.biomac.5b00549
