Influence of silk–silica fusion protein design on silica condensation in vitro and cellular calcification

Plowright, R, Dinjaski, N, Zhou, S, Belton, DJ, Kaplan, DL and Perry, CC ORCID: 0000-0003-1517-468X, 2016. Influence of silk–silica fusion protein design on silica condensation in vitro and cellular calcification. RSC Advances, 6 (26), pp. 21776-21788. ISSN 2046-2069

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Abstract

Biomaterial design via genetic engineering can be utilized for the rational functionalization of proteins to promote biomaterials integration and tissue regeneration. Spider silk has been extensively studied for its biocompatibility, biodegradability and extraordinary material properties. As a protein-based biomaterial, recombinant DNA derived derivatives of spider silks have been modified with biomineralization domains which lead to silica deposition and potentially accelerated bone regeneration. However, the influence of the location of the R5 (SSKKSGSYSGSKGSKRRIL) silicifying domain fused with the spider silk protein sequence on biosilicification process remains to be determined. Here we designed two silk-R5 fusion proteins that differed in the location of the R5 peptide, C- vs. N-terminus, where the spider silk domain consisted of a 15mer repeat of a 33 amino acid consensus sequence of the major ampullate dragline Spidroin 1 from Nephila clavipes (SGRGGLGG QG AGAAAAAGGA GQGGYGGLGSQGT). The chemical, physical and silica deposition properties of these recombinant proteins were assessed and compared to a silk 15mer control without the R5 present. The location of the R5 peptide did not have significant effect on wettability and surface energies, while the C - terminal location of the R5 promoted more controlled silica precipitation, suggesting differences in protein folding and possibly different access to charged amino acids that drive the silicification process. Further, cell compatibility in vitro, as well as the ability to promote human bone marrow derived mesenchymal stem cells (hMSC) differentiation were demonstrated for both variants of the fusion proteins.

Item Type: Journal article
Publication Title: RSC Advances
Creators: Plowright, R., Dinjaski, N., Zhou, S., Belton, D.J., Kaplan, D.L. and Perry, C.C.
Publisher: Royal Society of Chemistry
Date: 2016
Volume: 6
Number: 26
ISSN: 2046-2069
Identifiers:
NumberType
10.1039/C6RA03706BDOI
Divisions: Schools > School of Science and Technology
Depositing User: Linda Sullivan
Date Added: 24 Feb 2016 13:45
Last Modified: 09 Jun 2017 13:59
URI: http://irep.ntu.ac.uk/id/eprint/27029

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