Issue 3, 2012
From the journal:

Soft Matter

Characterization of a Ca binding-amphipathic silk-like protein and peptide with the sequence (Glu)8(Ala-Gly-Ser-Gly-Ala-Gly)4 with potential for bone repair

Aya Nagano,a   Hirohiko Sato,b   Yumi Tanioka,c   Yasumoto Nakazawa,c   David Knightd  and  Tetsuo Asakura*c  

* Corresponding authors

a Research and Technology Planning Office, Japan Medical Materials Corporation, Osaka, Japan

b Analysis Research Department, Chemical Laboratories, Nissan Chemical Industries LTD, Funabashi, Chiba, Japan

c Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan
E-mail: asakura@cc.tuat.ac.jp

d Oxford Biomaterials Ltd, Magdalen Centre, Oxford, UK

Abstract

Bombyx mori silk fibroin with the main sequence (Ala-Gly-Ser-Gly-Ala-Gly)n is a promising scaffold for bone regeneration not only on account of its excellent mechanical property as a structural matrix, but also for its slow biodegradability with adequate control of hydroxyapatite (HAP) deposition. Seeking to develop a material that might stimulate bone regeneration, we prepared a recombinant calcium binding-amphipathic silk-like protein [(Glu)8(Ala-Gly-Ser-Gly-Ala-Gly)4]4 by expression in E. coli. We also prepared the homologous peptide (Glu)8(Ala-Gly-Ser-Gly-Ala-Gly)4 by solid phase synthesis. The poly-L-glutamic acid was introduced into both protein and peptide because this sequence is involved in HAP-nucleating domains of bone sialoprotein. The recombinant protein was shown to bind relatively large quantities of Ca2+ ions in solution by a spectrophotometric assay and in the solid state by X-ray photoelectron spectroscopy. Changes in the electronic structure and local conformation of the peptide resulting from Ca2+ binding were studied using 13C solution NMR, especially 13C chemical shifts. We obtained evidence that Ca2+ bound to the poly-L-glutamic acid domains but not to the predominantly hydrophobic (Ala-Gly-Ser-Gly-Ala-Gly)4 domains. A remarkable conformational change induced by adsorption of the synthetic peptide on the HAP surface was also demonstrated using 13C solid state NMR.

Graphical abstract: Characterization of a Ca binding-amphipathic silk-like protein and peptide with the sequence (Glu)8(Ala-Gly-Ser-Gly-Ala-Gly)4 with potential for bone repair

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Article information

DOI
https://doi.org/10.1039/C1SM06646C
Article type
Paper
Submitted
29 Aug 2011
Accepted
11 Oct 2011
First published
07 Nov 2011

Soft Matter, 2012,8, 741-748

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Characterization of a Ca binding-amphipathic silk-like protein and peptide with the sequence (Glu)8(Ala-Gly-Ser-Gly-Ala-Gly)4 with potential for bone repair

A. Nagano, H. Sato, Y. Tanioka, Y. Nakazawa, D. Knight and T. Asakura, Soft Matter, 2012, 8, 741 DOI: 10.1039/C1SM06646C

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