Microphase Separation Controlled β-Sheet Crystallization Kinetics in Fibrous Proteins

Silk is a naturally occurring fibrous protein with a multiblock chain architecture. As Such, it has many similarities with synthetic block copolymers, including the possibility for beta-sheet crystallization restricted within the crystallizable blocks. The mechanism of isothermal crystallization kinetics of beta-sheet crystals in silk multiblock fibrous protein, is reported in this study. Kinetics theories, such as Avrami analysis which was established for studies of synthetic polymer crystal growth, are for the first time extended to investigate protein self-assembly in beta-sheet rich Bombyx mori silk fibroin samples, using time-resolved Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and synchrotron real-time wide-angle X-ray scattering (WAXS). The Avrami exponent, it, was close to 2 for all methods and crystallization temperatures, indicating formation of beta-sheet crystals in silk proteins is different from the 3-D spherulitic crystal growth found in synthetic polymers. Observations by scanning electron microscopy Support the view that the protein structures vary during the different stages of crystal growth, and show a microphase separation pattern after chymotrypsin enzyme biodegradation. We present a model to explain the crystallization of the multiblock silk fibroin protein, by analogy to block copolymers: crystallization of beta-sheets occurs under conditions of geometrical restriction caused by phase separation of the crystallizable and uncrystallizable blocks. This crystallization model could be widely applicable in other proteins with multiblock (i.e., crystallizable and noncrystallizable) domains.