Role of polyalanine domains in beta-sheet formation in spider silk block copolymers.

Genetically engineered spider silk-like block copolymers were studied to determine the influence of polyalanine domain size on secondary structure. The role of polyalanine block distribution on beta-sheet formation was explored using FT-IR and WAXS. The number of polyalanine blocks had a direct effect on the formation of crystalline beta-sheets, reflected in the change in crystallinity index as the blocks of polyalanines increased. WAXS analysis confirmed the crystalline nature of the sample with the largest number of polyalanine blocks. This approach provides a platform for further exploration of the role of specific amino acid chemistries in regulating the assembly of beta-sheet secondary structures, leading to options to regulate material properties through manipulation of this key component in spider silks.

[1]  T. Measey,et al.  The alanine-rich XAO peptide adopts a heterogeneous population, including turn-like and polyproline II conformations , 2007, Proceedings of the National Academy of Sciences.

[2]  Robert A. Grothe,et al.  Structure of the cross-β spine of amyloid-like fibrils , 2005, Nature.

[3]  R. E. Marsh,et al.  An investigation of the structure of silk fibroin. , 1955, Biochimica et biophysica acta.

[4]  D. Porter,et al.  Spider silk as a model biomaterial , 2006 .

[5]  Fritz Vollrath,et al.  Spider Silk Proteins – Mechanical Property and Gene Sequence , 2005, Zoological science.

[6]  T. M. Parker,et al.  Biocompatibility of the Bioelastic Materials, Poly(GVGVP) and Its γ-Irradiation Cross-Linked Matrix: Summary of Generic Biological Test Results , 1991 .

[7]  J. Kopeček,et al.  Genetically Engineered Block Copolymers: Influence of the Length and Structure of the Coiled-Coil Blocks on Hydrogel Self-Assembly , 2008, Pharmaceutical Research.

[8]  F. Kremer,et al.  Structure-property relationships in major ampullate spider silk as deduced from polarized FTIR spectroscopy , 2007, The European physical journal. E, Soft matter.

[9]  D. Kaplan,et al.  The effect of genetically engineered spider silk-dentin matrix protein 1 chimeric protein on hydroxyapatite nucleation. , 2007, Biomaterials.

[10]  N. Nevskaya,et al.  Infrared spectra and resonance interaction of amide‐I vibration of the antiparallel‐chain pleated sheet , 1976, Biopolymers.

[11]  S. Rammensee,et al.  Assembly mechanism of recombinant spider silk proteins , 2008, Proceedings of the National Academy of Sciences.

[12]  J C M van Hest,et al.  Elastin as a biomaterial for tissue engineering. , 2007, Biomaterials.

[13]  Maurille J. Fournier,et al.  Poly(L-alanylglycine) : Multigram-scale biosynthesis, crystallization, and structural analysis of chain-folded lamellae , 1997 .

[14]  D. Sogah,et al.  Self-assembly of beta-sheets into nanostructures by poly(alanine) segments incorporated in multiblock copolymers inspired by spider silk. , 2001, Journal of the American Chemical Society.

[15]  David L Kaplan,et al.  Collagen Structural Hierarchy and Susceptibility to Degradation by Ultraviolet Radiation. , 2008, Materials science & engineering. C, Materials for biological applications.

[16]  Alexander Kros,et al.  Noncovalent triblock copolymers based on a coiled-coil peptide motif. , 2008, Journal of the American Chemical Society.

[17]  A. Liwo,et al.  Further evidence for the absence of polyproline II stretch in the XAO peptide. , 2007, Biophysical journal.

[18]  T. Vuocolo,et al.  Synthesis and properties of crosslinked recombinant pro-resilin , 2005, Nature.

[19]  G. Freddi,et al.  Raman spectroscopic studies of silk fibroin fromBombyx mori , 1998 .

[20]  S. Funari,et al.  The Effect of PEG Crystallization on the Morphology of PEG/Peptide Block Copolymers Containing Amyloid β‐Peptide Fragments , 2008 .

[21]  David L Kaplan,et al.  Spider silks and their applications. , 2008, Trends in biotechnology.

[22]  Randolph V Lewis,et al.  Spider silk: ancient ideas for new biomaterials. , 2006, Chemical reviews.

[23]  David L. Kaplan,et al.  Dynamic Protein−Water Relationships during β-Sheet Formation , 2008 .

[24]  Pawel Sikorski,et al.  Molecular basis for amyloid fibril formation and stability. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[25]  D. Kaplan,et al.  Conformational transitions in model silk peptides. , 2000, Biophysical journal.

[26]  Lawrence F. Drummy,et al.  Correlation of the β-sheet crystal size in silk fibers with the protein amino acid sequence. , 2007, Soft matter.

[27]  H M Berman,et al.  Hydration structure of a collagen peptide. , 1995, Structure.

[28]  S. Lee,et al.  Solution behavior of synthetic silk peptides and modified recombinant silk proteins , 2006 .

[29]  R. Lewis,et al.  Hypotheses that correlate the sequence, structure, and mechanical properties of spider silk proteins. , 1999, International journal of biological macromolecules.

[30]  L. Serpell,et al.  Spider silk and amyloid fibrils: a structural comparison. , 2007, Macromolecular bioscience.

[31]  F. Vollrath,et al.  Biological liquid crystal elastomers. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[32]  G. Szulgit The echinoderm collagen fibril: a hero in the connective tissue research of the 1990s , 2007, BioEssays : news and reviews in molecular, cellular and developmental biology.

[33]  D. Kaplan,et al.  Mechanisms of silk fibroin sol-gel transitions. , 2006, The journal of physical chemistry. B.

[34]  David T. Grubb,et al.  Fiber Morphology of Spider Silk: The Effects of Tensile Deformation , 1997 .

[35]  B. Lotz,et al.  Twisted single crystals of Bombyx mori silk fibroin and related model polypeptides with beta structure. A correlation with the twist of the beta sheets in globular proteins. , 1982, Journal of molecular biology.

[36]  David L Kaplan,et al.  RGD-functionalized bioengineered spider dragline silk biomaterial. , 2006, Biomacromolecules.

[37]  David L Kaplan,et al.  Silk: molecular organization and control of assembly. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[38]  David L. Kaplan,et al.  Mechanism of silk processing in insects and spiders , 2003, Nature.

[39]  T. Scheibel Protein fibers as performance proteins: new technologies and applications. , 2005, Current opinion in biotechnology.

[40]  Z. Shao,et al.  Analysis of spider silk in native and supercontracted states using Raman spectroscopy , 1999 .

[41]  J. Gosline,et al.  The mechanical design of spider silks: from fibroin sequence to mechanical function. , 1999, The Journal of experimental biology.

[42]  Derek N Woolfson,et al.  Peptide-based fibrous biomaterials: Some things old, new and borrowed. , 2006, Current opinion in chemical biology.

[43]  Andrea Lomander,et al.  Hierarchical self-assembly of a coiled-coil peptide into fractal structure. , 2005, Nano letters.

[44]  C. Viney,et al.  Non-periodic lattice crystals in the hierarchical microstructure of spider (major ampullate) silk. , 1997, Biopolymers.

[45]  G. Jeschke,et al.  Structure and dynamics of self-assembled poly(ethylene glycol) based coiled-coil nano-objects. , 2004, ChemPhysChem.

[46]  D. Kaplan,et al.  Controlling beta-sheet assembly in genetically engineered silk by enzymatic phosphorylation/dephosphorylation. , 2000, Biochemistry.

[47]  H. Edwards,et al.  Raman spectroscopic studies of silk , 1995 .

[48]  David L Kaplan,et al.  Self-assembly of genetically engineered spider silk block copolymers. , 2009, Biomacromolecules.

[49]  D. Downing,et al.  β-Helical Fibrils from a Model Peptide , 1997 .

[50]  David L Kaplan,et al.  Mapping domain structures in silks from insects and spiders related to protein assembly. , 2004, Journal of molecular biology.

[51]  R. Fraser,et al.  Poly-l-alanylglycyl-l-alanylglycyl-l-serylglycine: a model for the crystalline regions of silk fibroin. , 1966, Journal of molecular biology.

[52]  M. E. Demont,et al.  Spider silk as rubber , 1984, Nature.

[53]  Thierry Lefèvre,et al.  Protein secondary structure and orientation in silk as revealed by Raman spectromicroscopy. , 2007, Biophysical journal.

[54]  Kang Chen,et al.  Conformation of the backbone in unfolded proteins. , 2006, Chemical reviews.

[55]  K. Kohler,et al.  Molecular mechanisms of spider silk , 2006, Cellular and Molecular Life Sciences CMLS.

[56]  A. Barth Infrared spectroscopy of proteins. , 2007, Biochimica et biophysica acta.

[57]  P. Garside,et al.  Characterization of Historic Silk by Polarized Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy for Informed Conservation , 2005, Applied spectroscopy.

[58]  I. Hamley,et al.  Solution self-assembly of hybrid block copolymers containing poly(ethylene glycol) and amphiphilic beta-strand peptide sequences. , 2005, Biomacromolecules.

[59]  E. Bramanti,et al.  Solid state (13)C NMR and FT-IR spectroscopy of the cocoon silk of two common spiders. , 2005, Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy.

[60]  C. Riekel,et al.  Aspects of X-ray diffraction on single spider fibers. , 1999, International journal of biological macromolecules.

[61]  David L. Kaplan,et al.  Determining Beta-Sheet Crystallinity in Fibrous Proteins by Thermal Analysis and Infrared Spectroscopy , 2006 .

[62]  K. Beck,et al.  Supercoiled Protein Motifs: The Collagen Triple-Helix and the α-Helical Coiled Coil , 1998 .

[63]  Tetsuo Asakura,et al.  Conformational characterization of Bombyx mori silk fibroin in the solid state by high-frequency carbon-13 cross polarization-magic angle spinning NMR, x-ray diffraction, and infrared spectroscopy , 1985 .