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Triggered Morphology Generation in a Biosynthetic Model Spider Dragline Silk Protein

Published online by Cambridge University Press:  02 July 2020

R. Valluzzi ,
S. Szela ,
D. Kirschner  and
D. Kaplan
R. Valluzzi
Affiliation:
Department of Chemical Engineering, Biotechnology Center, Tufts University, Medford, MA02155
S. Szela
Affiliation:
Department of Chemical Engineering, Biotechnology Center, Tufts University, Medford, MA02155
D. Kirschner
Affiliation:
Department of Biology, Boston College, Allston, MA02467
D. Kaplan
Affiliation:
Department of Chemical Engineering, Biotechnology Center, Tufts University, Medford, MA02155
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Extract

Recombinant DNA techniques were used to prepare a protein modeled after the consensus sequence of Nephila clavipesspider dragline silk, incorporating methionine residues to serve as redox “triggers”. In addition a water-soluble 27 residue peptide model of the dragline silk consensus amorphous sequence, representing a single amorphous block in the protein sequence, was prepared and characterized to gain additional insight into the behavior of the amorphous phase. X-ray diffraction, electron diffraction, transmission electron microscopy (TEM), and Fourier transform infrared spectroscopy (FTIR) were used to characterize the ability of the recombinant protein to form (β-sheet crystals and the effect of the oxidation state of the redox trigger on crystallinity and noncrystalline order in the sample. The formation of intractable β-sheet crystallites is a major cause of insolubility in proteins that can form this type of secondary structure. Changes in crystallinity were observed when triggered/reduced (insoluble) and untriggered/oxidized (soluble) protein samples were compared.

Type
Biopolymers and Biomemetics
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1214 - 1215
Copyright
Copyright © Microscopy Society of America

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References

l)Winkler, S.; Szela, S.; Avtges, P.; Valluzzi, R.; Kerschner, D.; Kaplan, D. L.Int. J. Biol. Mol. 1998Google Scholar
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5)We would like to acknowledge support from the National Science Foundation; NSF DMR- 9708062 and NSF BES-9727401, the NSF MRSEC program, and from the W. M. Keck Foundation Biomimetic Materials and Polymer Morphology LaboratoriesGoogle Scholar