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Polymers 2018, 10(11), 1193; https://doi.org/10.3390/polym10111193

An RGD-Containing Peptide Derived from Wild Silkworm Silk Fibroin Promotes Cell Adhesion and Spreading

1
National Engineering Laboratory for Modern Silk, College of Textile and Clothing Engineering, Soochow University, No. 199 Ren-ai Road, Suzhou Industrial Park, Suzhou 215123, China
2
Department of Biological Sciences, Xi’an Jiaotong-Liverpool University, No. 111 Ren-ai Road, Suzhou Industrial Park, Suzhou 215123, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 10 October 2018 / Revised: 22 October 2018 / Accepted: 24 October 2018 / Published: 26 October 2018
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Abstract

Arginine-Glycine-Aspartate (RGD) tripeptide can promote cell adhesion when present in the amino acid of proteins such as fibronectin. In order to demonstrate the bioactivity of an RGD-containing silk protein, a gene encoding the RGD motif-containing peptide GSGAGGRGDGGYGSGSS (–RGD–) derived from nonmulberry silk was designed and cloned, then multimerised and inserted into a commercial pGEX expression vector for recombinant expression of (–RGD–)n peptides. Herein, we focus on two glutathione-S-transferase (GST)-tagged fusion proteins, GST–(–RGD–)4 and GST–(–RGD–)8, which were expressed in Escherichia coli BL21, purified by GST affinity chromatography, and analyzed with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). Target peptides (–RGD–)4 and (–RGD–)8 (6.03 and 11.5 kDa) were cleaved from the GST-tag by thrombin digestion, as verified with MS and SDS-PAGE. Isoelectric point analysis confirmed that target peptides were expressed and released in accordance with the original design. Target peptides self-assembled into a mainly α-helical structure, as determined by circular dichroism spectroscopy. Furthermore, (–RGD–)4 and (–RGD–)8 modified mulberry silk fibroin films were more effective for rapid cell adhesion, spreading and proliferative activity of L929 cells than some chemically synthesized RGD peptides modified and mulberry silk lacking the RGD motif. View Full-Text
Keywords: wild silkworm; silk fibroin; RGD peptides; biosynthesis; cell adhesion; cell proliferation wild silkworm; silk fibroin; RGD peptides; biosynthesis; cell adhesion; cell proliferation
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Kang, Z.; Wang, Y.; Xu, J.; Song, G.; Ding, M.; Zhao, H.; Wang, J. An RGD-Containing Peptide Derived from Wild Silkworm Silk Fibroin Promotes Cell Adhesion and Spreading. Polymers 2018, 10, 1193.

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