Characterization by Raman microspectroscopy of the strain-induced conformational transition in fibroin fibers from the silkworm Samia cynthia ricini.
Raman microspectroscopy has been used to quantitatively study the effect of a mechanical deformation on the conformation and orientation of Samia cynthia ricini (S. c. ricini) silk fibroin. Samples were obtained from the aqueous solution stored in the silk gland and stretched at draw ratios (lambda) ranging from 0 to 11. Using an appropriate band decomposition procedure, polarized and orientation-insensitive spectra have been analyzed to determine order parameters and the content of secondary structures, respectively. The data unambiguously show that, in response to mechanical deformation, S. c. ricini fibroin undergoes a cooperative alpha-helix to beta-sheet conformational transition above a critical draw ratio of 4. The alpha-helix content decreases from 33 to 13% when lambda increases from 0 to 11, while the amount of beta-sheets increases from 15 to 37%. In comparison, cocoon silk is devoid of alpha-helical structure and always contains a larger amount of beta-sheets. Although the presence of isosbestic points in different spectral regions reveals that the conformational change induced by mechanical deformation is a two-state process, our results suggest that part of the glycine residues might be incorporated into beta-poly(alanine) structures. The beta-sheets are initially isotropically distributed and orient along the fiber axis as lambda increases, but do not reach the high level of orientation found in the cocoon fiber. The increase in the orientation level of the beta-sheets is found to be concomitant with the alpha --> beta conformational conversion, whereas alpha-helices do not orient under the applied strain but are rather readily converted into beta-sheets. The components assigned to turns exhibit a small orientation perpendicular to the fiber axis in stretched samples, showing that, overall, the polypeptide chains are aligned along the stretching direction. Our results suggest that, in nature, factors other than stretching contribute to the optimization of the amount of beta-sheets and the high degree of orientation found in natural cocoon silk.
Rousseau
Marie-Eve
ME
Département de Chimie et CERSIM, Université Laval, Québec, QC, Canada G1K 7P4.
Beaulieu
Lilyane
L
Lefèvre
Thierry
T
Paradis
Joanie
J
Asakura
Tetsuo
T
Pézolet
Michel
M
D000465:Algorithms; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D012831:Bombyx; D005349:Fibroins / Q000737:chemistry*; D046911:Macromolecular Substances; D008422:Materials Testing; D008968:Molecular Conformation; D010455:Peptides; D015202:Protein Engineering; D017433:Protein Structure, Secondary; D047011:Silk / Q000378:metabolism; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013314:Stress, Mechanical
D001672:Biocompatible Materials; D046911:Macromolecular Substances; D010455:Peptides; D047011:Silk; D005349:Fibroins