Molecular Dynamics of Spider Dragline Silk Fiber Investigated by ²H MAS NMR

The molecular dynamics of the proteins that comprise spider dragline silk were investigated with solid-state ²H magic angle spinning (MAS) NMR line shape and spin–lattice relaxation time (T₁) analysis. The experiments were performed on ²H/¹³C/¹⁵N-enriched N. clavipes dragline silk fibers. The silk protein side-chain and backbone dynamics were probed for Ala-rich regions (β-sheet and 3₁-helical domains) in both native (dry) and supercontracted (wet) spider silk. In native (dry) silk fibers, the side chains in all Ala containing regions undergo similar fast methyl rotations (>10⁹ s^–1), while the backbone remains essentially static (<10² s^–1). When the silk is wet and supercontracted, the presence of water initiates fast side-chain and backbone motions for a fraction of the β-sheet region and 3₁-helicies. β-Sheet subregion 1 ascribed to the poly­(Ala) core exhibits slower dynamics, while β-sheet subregion 2 present in the interfacial, primarily poly­(Gly-Ala) region that links the β-sheets to disordered 3₁-helical motifs, exhibits faster motions when the silk is supercontracted. Particularly notable is the observation of microsecond backbone motions for β-sheet subregion 2 and 3₁-helicies. It is proposed that these microsecond backbone motions lead to hydrogen-bond disruption in β-sheet subregion 2 and helps to explain the decrease in silk stiffness when the silk is wet and supercontracted. In addition, water mobilizes and softens 3₁-helical motifs, contributing to the increased extensibility observed when the silk is in a supercontracted state. The present study provides critical insight into the supercontraction mechanism and corresponding changes in mechanical properties observed for spider dragline silks.