at Google ScholarTitle data
Stengel, Dillan ; Saric, Merisa ; Johnson, Hannah R. ; Schiller, Tim ; Diehl, Johannes ; Chalek, Kevin ; Onofrei, David ; Scheibel, Thomas ; Holland, Gregory P.:
Tyrosine's Unique Role in the Hierarchical Assembly of Recombinant Spider Silk Proteins : From Spinning Dope to Fibers.
In: Biomacromolecules.
Vol. 24
(2023)
Issue 3
.
- pp. 1463-1474.
ISSN 1526-4602
DOI: https://doi.org/10.1021/acs.biomac.2c01467
Abstract in another language
Producing recombinant spider silk fibers that exhibit mechanical properties approaching native spider silk is highly dependent on the constitution of the spinning dope. Previously published work has shown that recombinant spider silk fibers spun from dopes with phosphate-induced pre-assembly (biomimetic dopes) display a toughness approaching native spider silks far exceeding the mechanical properties of fibers spun from dopes without pre-assembly (classical dopes). Dynamic light scattering experiments comparing the two dopes reveal that biomimetic dope displays a systematic increase in assembly size over time, while light microscopy indicates liquid–liquid-phase separation (LLPS) as evidenced by the formation of micron-scale liquid droplets. Solution nuclear magnetic resonance (NMR) shows that the structural state in classical and biomimetic dopes displays a general random coil conformation in both cases; however, some subtle but distinct differences are observed, including a more ordered state for the biomimetic dope and small chemical shift perturbations indicating differences in hydrogen bonding of the protein in the different dopes with notable changes occurring for Tyr residues. Solid-state NMR demonstrates that the final wet-spun fibers from the two dopes display no structural differences of the poly(Ala) stretches, but biomimetic fibers display a significant difference in Tyr ring packing in non-β-sheet, disordered helical domains that can be traced back to differences in dope preparations. It is concluded that phosphate pre-orders the recombinant silk protein in biomimetic dopes resulting in LLPS and fibers that exhibit vastly improved toughness that could be due to aromatic ring packing differences in non-β-sheet domains that contain Tyr.